Dorper Sheep Research Articles

Molecular and genealogical analyses reveal multiple sources of the mutation associated with dermatosparaxis in Brazilian White Dorper sheep

Dermatosparaxis, also known as Ehlers-Danlos syndrome, is a recessive genetic disorder characterized by defects in collagen production, leading to skin fragility and delayed wound healing. In sheep this disease is associated to a transversion c.421G>T located on exon 2 of the gene ADAMTS2 and presents a special importance for the White Dorper breed. This study aims to identify carriers for the mutation associated with dermatosparaxis in Brazilian White Dorper herds and also to better understand the origin and the possibility of dissemination of this disease combining molecular and genealogical approaches. Genotyping by PCR-RFLP revealed the presence of seven out 66 heterozygotes, resulting in an estimated frequency of 5.3% for the dermatosparaxis associated allele in Brazilian White Dorper sheep. Pedigree analysis permitted the identification of two separate groups of individuals carrying the dermatosparaxis associated allele, one represented by descendants from individuals imported from South Africa and the second originated from animals imported from Australia. These findings suggest that the implementation of a program aiming to eradicate/control of dermatosparaxis in Brazilian White Dorper sheep is needed and the success of this strategy initially depends on the utilization of a molecular testing to detect carriers.

Examination of microsatellite markers of Dorper sheep breed

Number of not woolly and molty sheep exceeds 60 million throughout the world. Their numbers and their importance is growing, still they have appeared in the past two decades all over in North-America, Australia, New-Zealand and also in Europe. The South African Dorper has been a pioneer among them in Hungary. It was introduced in 2006 in the country. The Dorper sheep is the second largest breed in South Africa, which was developed from the crossing of Dorset Horn and the Blackhead Persian. The aim of the EU Member States in terms of this specific breed is increasing the small populations, improving the productive qualities, in addition to this avoiding inbreeding. However, finding appropriate breeding stock is difficult due to the small size of available populations and also to the suspected common of origin. With the help of various molecular genetic methods we could get a total view of the genetic background of these flocks. Nowadays the most commonly known and used genetic markers are microsatellites, because their applications give fast, accurate and easily reproducible results. There is no specific descriptive information on the genetic background of Dorper populations in the various EU countries , also regarding diversity between populations. Therefore in our work we want to optimize the conditions of applicability of 31 selected microsatellite reactions as a first step of mapping the entire genetic background of the different EU Dorper populations.

Determinação de valores de referência séricos para os eletrólitos magnésio, cloretos, cálcio e fósforo em ovinos das raças Dorper e Santa Inês

Resumo: A ovinocultura no Brasil é uma atividade em grande expansão e, com o aumento da demanda mundial por carne ovina, aumentou-se o interesse no monitoramento da sanidade do rebanho, utilizando diversas ferramentas como auxiliares no diagnóstico clínico, tais como os intervalos de referência séricos. Os elementos minerais constituem 2 a 5,5% do corpo dos vertebrados, exercendo diversas funções no organismo. O objetivo deste trabalho foi obter intervalos de referência para os eletrólitos magnésio, fósforo, cloreto e cálcio para ovinos das raças Dorper e Santa Inês. Foram coletados soros de 487 animais clinicamente sadios, sendo 146 da raça Dorper e 341 da raça Santa Inês. Os eletrólitos foram mensurados utilizando-se kits comerciais. Os dados foram analisados quanto à raça, sexo e idade, e os intervalos de referência determinados. Os resultados revelaram diferenças significativas nos intervalos de referência obtidos para os eletrólitos cálcio e magnésio na variável raça, e para o eletrólito fósforo na variável faixa etária e, quando confrontados com valores de referência já publicados, comprovou-se a existência de diferença estatística significativa entre os mesmos em todos os analitos estudados.

ASPIRAÇÃO FOLICULAR VIDEOLAPAROSCÓPICA COMPARATIVA EM OVELHAS DORPER E SANTA INÊS

Resumo Este estudo teve por objetivo comparar o número de folículos aspirados, oócitos recuperados, qualidade e viabilidade oocitária entre ovelhas das raças Dorper e Santa Inês, submetidas à aspiração folicular por vídeolaparoscopia. Para realização deste estudo, foram efetuadas 118 aspirações foliculares, das quais 65 foram realizadas em ovelhas da raça Dorper e 53 em ovelhas da raça Santa Inês. Os resultados encontrados para as ovelhas Dorper foram 28,31 ± 17,42 folículos/ovelha, 12,21 ± 6,74 oócitos/ovelha, taxa de recuperação de 47,9% e 8,93 ± 5,25 oócitos viáveis/ovelha. Para as ovelhas Santa Inês foram, 22,14 ± 16,08 folículos/ovelha, 12,64 ± 8,29 oócitos/ovelha, taxa de recuperação de 55% e 8,92 ± 6,58 oócitos viáveis/ovelha. Não houve influência das raças sobre o número de folículos aspirados, oócitos recuperados, taxa de recuperação, qualidade e viabilidade oocitária em aspiração folicular por vídeolaparoscopia.

Prevalence study of SNP c.421G>T in the ADAMTS2 gene responsible for dermatosparaxis in White Dorper sheep in Brazil

Abstract: Dermatosparaxis is an autosomal recessive disorder of connective tissue; the disorder is clinically characterized by skin fragility and hyperextensibility. Dermatosparaxis in White Dorper sheep is caused by a single nucleotide polymorphism (SNP) (c.421G>T) in the ADAM metalloproteinase with thrombospondin type 1 motif, 2 (ADAMTS2) gene. The aim of this study was to investigate the prevalence of this SNP in a White Dorper herd in São Paulo state, Brazil. In this study, we collected blood DNA samples from 303 White Dorper sheep and performed polymerase chain reaction to amplify the SNP region. The samples were sequenced to determine the presence of the SNP in the ADAMTS2 gene. The SNP prevalence in the studied population was 15.5%; this finding indicates that more effective control measures should be used to prevent the inheritance of SNP c.421G>T in the ADAMTS2 gene in Brazilian White Dorper herds.

Deterioration of teeth and alveolar bone loss due to chronic environmental high-level fluoride and low calcium exposure.

Health risks due to chronic exposure to highly fluoridated groundwater could be underestimated because fluoride might not only influence the teeth in an aesthetic manner but also seems to led to dentoalveolar structure changes. Therefore, we studied the tooth and alveolar bone structures of Dorper sheep chronically exposed to very highly fluoridated and low calcium groundwater in the Kalahari Desert in comparison to controls consuming groundwater with low fluoride and normal calcium levels within the World Health Organization (WHO) recommended range. Two flocks of Dorper ewes in Namibia were studied. Chemical analyses of water, blood and urine were performed. Mineralized tissue investigations included radiography, HR-pQCT analyses, histomorphometry, energy-dispersive X-ray spectroscopy and X-ray diffraction-analyses. Fluoride levels were significantly elevated in water, blood and urine samples in the Kalahari group compared to the low fluoride control samples. In addition to high fluoride, low calcium levels were detected in the Kalahari water. Tooth height and mandibular bone quality were significantly decreased in sheep, exposed to very high levels of fluoride and low levels of calcium in drinking water. Particularly, bone volume and cortical thickness of the mandibular bone were significantly reduced in these sheep. The current study suggests that chronic environmental fluoride exposure with levels above the recommended limits in combination with low calcium uptake can cause significant attrition of teeth and a significant impaired mandibular bone quality. In the presence of high fluoride and low calcium-associated dental changes, deterioration of the mandibular bone and a potential alveolar bone loss needs to be considered regardless whether other signs of systemic skeletal fluorosis are observed or not.

Molecular cloning, characterization and tissue specificity of the expression of the ovine CSRP2 and CSRP3 genes from Small-tail Han sheep (Ovis aries)

Two genes, cysteine- and glycine-rich protein 2 (CSRP2) and cysteine- and glycine-rich protein 3 (CSRP3), play important roles in tissue-specific cell growth and development. However, few CSRP2 and CSRP3 genes have been functionally characterized in sheep. In this study, the full-length cDNAs of the CSRP2 and CSRP3 genes were cloned from Small-tail Han sheep by rapid amplification of cDNA ends-PCR. The GenBank accession numbers of the full-length CSRP2 and CSRP3 cDNA sequences are KJ743957 and KJ743958, respectively. The full-length cDNA of ovine CSRP2 was 917bp, with a 582-bp open reading frame encoding 193 amino acids. The complete ovine CSRP3 cDNA was 917bp, with a 585-bp open reading frame encoding 194 amino acids. Alignment and phylogenetic analyses revealed that their amino acid sequences are highly similar to those of other vertebrates, all of which contain two conserved LIM-only domains and a relatively conserved nuclear targeting sequence. To further validate the functions of the two genes, their mRNA expression patterns were evaluated in various Small-tail Han and Dorper sheep tissues using qRT-PCR analyses. CSRP2 was mainly detected in the aorta, whereas CSRP3 was highly concentrated in the heart and the muscle. CSRP3 was expressed to a higher level in the hearts of Small-tail Han sheep than in Dorper sheep (P<0.05). However, the opposite was found in the muscle (the longissimus dorsi and biceps femoris); CSRP3 was expressed to a higher level in Dorper sheep than in Small-tail Han sheep (P<0.05). We quantified the CRP3 protein (coded by the CSRP3 gene) levels in different tissues in Small-tail Han and Dorper sheep. We also detected a putative isoform of the CRP3 protein in sheep, which was significantly different in the heart tissue of the two breeds (P<0.05). The expression patterns of the two genes' mRNAs and CRP3 protein showed clear tissue specificity in both sheep breeds.

A genome scan of recent positive selection signatures in three sheep populations

Domesticated sheep have been exposed to artificial selection for the production of fiber, meat, and milk as well as to natural selection. Such selections are likely to have imposed distinctive selection signatures on the sheep genome. Therefore, detecting selection signatures across the genome may help elucidate mechanisms of selection and pinpoint candidate genes of interest for further investigation. Here, detection of selection signatures was conducted in three sheep breeds, Sunite (n=66), German Mutton (n=159), and Dorper (n=93), using the Illumina OvineSNP50 Genotyping BeadChip array. Each animal provided genotype information for 43 273 autosomal single nucleotide polymorphisms (SNPs). We adopted two complementary haplotype-based statistics of relative extended haplotype homozygosity (REHH) and the cross-population extended haplotype homozygosity (XP-EHH) tests. In total, 707, 755, and 438 genomic regions subjected to positive selection were identified in Sunite, German Mutton, and Dorper sheep, respectively, and 42 of these regions were detected using both REHH and XP-EHH analyses. These genomic regions harbored many important genes, which were enriched in gene ontology terms involved in muscle development, growth, and fat metabolism. Fourteen of these genomic regions overlapped with those identified in our previous genome-wide association studies, further indicating that these genes under positive selection may underlie growth developmental traits. These findings contribute to the identification of candidate genes of interest and aid in understanding the evolutionary and biological mechanisms for controlling complex traits in Chinese and western sheep.

Comparative daily energy expenditure and water turnover by Dorper and Merino sheep measured using doubly labelled water

Details of the energy (food) requirements of domestic herbivores are essential for predicting grazing pressures and subsequent ecological impacts on rangelands. However, these details are lacking for some of the more recently introduced sheep breeds to Australia, such as the Dorper breed sheep, which are principally meat sheep, and it is uncertain how they compare with the traditional Merino, a wool-breed, sheep. We used the doubly labelled water method to compare the field metabolic rate and water turnover rate of Dorpers and Merinos grazing together in a small holding paddock in a typical rangeland environment. We found no significant differences in field metabolic rate (Dorpers 481 ± 125 kJ and Merinos 500 ± 109 kJ kg–0.73 day–1) or water turnover rate (Dorpers 397 ± 57 mL and Merinos 428 ± 50 mL kg–0.8 day–1). As such we conclude that under controlled conditions with limited movement and ready access to feed and water, dry sheep equivalent of 1 is appropriate for Dorpers (that is, one Dorper ewe had a grazing requirement equal to one standard, dry Merino wether). However, we also found that the field metabolic rate for Merinos under these conditions was only around half that measured in published studies for animals ranging freely in a large paddock system. This suggests that more work is needed to fully appreciate the energetic and grazing impacts of Dorpers versus Merinos under more realistic grazing conditions (e.g. large paddock systems) where feed and water are more spread. It also highlights limitations of the current dry sheep equivalent rating system, which has been derived from laboratory measures of sheep metabolic rates.

Addition of sucrose on cryoprotectant solution of vitrification enhances the quality of Dorper sheep embryos produced in vivo

This study aimed to evaluate the efficacy of adding sucrose in vitrification solution of ovine embryos produced in vivo. Forty Dorper ewes were selected and superovulated. Immediately prior to the embryo collection by laparotomy, a laparoscopy was performed to verify the superovulatory response. The recovered flushing was followed by embryo evaluation and embryos were divided in two experimental groups where embryos from Control group were submitted to a traditional vitrification protocol and embryos from Sucrose group to a modified vitrification protocol with sucrose. After warming, embryos were again divided regarding cryoprotectant removal (Indirect) or not (Direct). The embryo quality was classified as embryos of degrees I (excellent or good), II (regular), III (poor) and IV (dead or degenerate). It was also verified the homogeneity of mass, occurrence of embryonic mass retraction and rupture of pellucid zone. The results were expressed as percentages and were subjected to Chi-square test with P &lt; 0.05. The embryos vitrified in the presence of sucrose had lower proportions of lower-quality embryos after warming (22.20 vs. 44.50%), higher percentages of homogeneous embryos after warming (63.89 vs. 38.89 %) while concerning other parameters there was no difference between these groups. It can be concluded that the addition of 0.4 M sucrose during vitrification improves the embryo quality.

Addition of sucrose on cryoprotectant solution of vitrification enhances the quality of Dorper sheep embryos produced in vivo

&lt;p&gt;This study aimed to evaluate the efficacy of adding sucrose in vitrification solution of ovine embryos produced &lt;em&gt;in vivo&lt;/em&gt;. Forty Dorper ewes were selected and superovulated. Immediately prior to the embryo collection by laparotomy, a laparoscopy was performed to verify the superovulatory response. The recovered flushing was followed by embryo evaluation and embryos were divided in two experimental groups where embryos from Control group were submitted to a traditional vitrification protocol and embryos from Sucrose group to a modified vitrification protocol with sucrose. After warming, embryos were again divided regarding cryoprotectant removal (Indirect) or not (Direct). The embryo quality was classified as embryos of degrees I (excellent or good), II (regular), III (poor) and IV (dead or degenerate). It was also verified the homogeneity of mass, occurrence of embryonic mass retraction and rupture of pellucid zone. The results were expressed as percentages and were subjected to Chi-square test with P &amp;lt; 0.05. The embryos vitrified in the presence of sucrose had lower proportions of lower-quality embryos after warming (22.20 vs. 44.50%), higher percentages of homogeneous embryos after warming (63.89 vs. 38.89 %) while concerning other parameters there was no difference between these groups. It can be concluded that the addition of 0.4 M sucrose during vitrification improves the embryo quality.&lt;/p&gt;

Carcass muscularity and adiposity of different sheep and goat genotypes

SUMMARY Carcass muscularity and adiposity of different goat and sheep genotypes grazing in the Caatinga rangeland were evaluated. A total of 40 uncastrated males, with an initial average age and body weight of 120 days and 18.76 kg, respectively, were used: 10 mixed breed goats (MB), 10 cross Boer goats (½ Boer x ½ MB), 10 MB sheep and 10 cross Dorper sheep (½ Dorper x ½ MB). The cross Dorper had higher mean scores for rib eye area, leg muscularity index, weight of total muscles of the leg and conformation compared to MB goats. There was no significant difference between genotypes for the meat texture. Meat color was darker for goat genotypes than for sheep genotypes. The quantitative aspects of carcass adiposity, renal fat, inguinal fat, and subcutaneous and total leg fat showed to be higher for sheep than for goat genotypes. Carcass finishing and leg compactness index were superior for the cross Dorper genotype. Carcass muscularity and adiposity were better in sheep than in goat genotypes, indicating that sheep have a greater muscle and fat tissue development, giving to them a greater potential for beef production.

Molecular cloning and mRNA expression analysis of sheep MYL3 and MYL4 genes

Using longissimus dorsi muscles of Dorper sheep as the experimental materials, the complete cDNAs of ovine MYL3 (Myosin light chain 3) and MYL4 (Myosin light chain 4) genes were cloned using RT-PCR, 5′ RACE and 3′ RACE. We obtained 925-bp and 869-bp full-length cDNAs and submitted their sequences to GenBank as accession numbers of KJ710703 and KJ768855, respectively. The cDNAs contained 600-bp and 582-bp open reading frames (ORFs) and encoded proteins comprising 199 and 193 amino acid residues, respectively. Neither protein was predicted to have a signal peptide, but both were predicted to have several N-glycosylation, O-glycosylation, and phosphorylation sites. The secondary structures of MYL3 and MYL4 were predicted to be 40.70% and 48.70% α- helical, respectively. Sequence alignment showed that the MYL3 and MYL4 proteins of Ovis aries both shared more than 91% amino acid sequence similarity with those of Mus musculus, Homo sapiens, Rattus norvegicus, Bos taurus, and Sus scrofa. The levels of MYL3 and MYL4 mRNA in various sheep tissues were determined using qRT-PCR. The results showed that both mRNAs were highly expressed in the heart. This study has established a foundation for further investigation of the ovine MYL3 and MYL4 genes.

Participatory definition of breeding objectives for sheep breeds under pastoral systems--the case of Red Maasai and Dorper sheep in Kenya.

Crossing local breeds with exotic breeds may be an option for increased livestock productivity. However, there is a risk for endangerment of the local breeds. One such case is in Kenya where the imported Dorper breed is used for crossbreeding with Red Maasai sheep. The aim of this study was to investigate farmers’ trait preferences as a basis for determination of breeding objectives for Red Maasai and Dorper sheep at two sites, Amboseli and Isinya, in Kenya. Within their own flock, each farmer identified three ewes representing the best, average and poorest within each breed group: Red Maasai, Dorper and Crosses. Farmers gave reasons for their ranking. Body measurements and weights were also taken. At the harshest site, Amboseli, differences between breed groups in body weight were small and breeds were equally preferred. In Isinya, where environmental conditions are better and farmers are more market oriented, Dorper and Crosses had significantly higher body weights and market prices and were thus preferred by the farmers. Red Maasai were preferred for their maternal and adaptive traits. Breeding objectives should emphasize growth traits and milk production in both breeds at both sites. Body condition needs to be specifically considered in the breeding objectives for sheep in Amboseli, whereas adaptive traits need to be generally emphasized in Dorper.

Detection of Selection Signatures on the X Chromosome in Three Sheep Breeds.

Artificial selection has played a critical role in animal breeding. Detection of artificial selection footprints in genomic regions can provide insights for understanding the function of specific phenotypic traits and better guide animal breeding. To more fully understand the relationship between genomic composition and phenotypic diversity arising from breed development, a genome-wide scan was conducted using an OvineSNP50 BeadChip and integrated haplotype score and fixation index analyses to detect selection signatures on the X chromosome in three sheep breeds. We identified 49, 34, and 55 candidate selection regions with lengths of 27.49, 16.47, and 25.42 Mb in German Mutton, Dorper, and Sunit sheep, respectively. Bioinformatics analysis showed that some of the genes in these regions with selection signatures, such as BMP15, were relevant to reproduction. We also identified some selection regions harboring genes that had human orthologs, including BKT, CENPI, GUCY2F, MSN, PCDH11X, PLP1, VSIG4, PAK3, WAS, PCDH19, PDHA1, and SRPX2. The VSIG4 and PCDH11X genes are associated with the immune system and disease, PDHA1 is associated with biosynthetic related pathways, and PCDH19 is expressed in the nervous system and skin. These genes may be useful as candidate genes for molecular breeding.

Fatty acid composition of the ovine longissimus dorsi muscle: effect of feed restriction in three breeds of different origin.

Muscle fatty acid profile reflects the body condition of animals and has a noticeable effect on meat quality. Herein, longissimus dorsi muscle of three different sheep breeds, Damara (a fat-tailed breed), Dorper and Australian Merino sheep, was analysed for fatty acid composition. The three breeds were subjected to two distinctive feeding levels (ad libitum and restricted feeding) over 42 days. The Damara sheep revealed several differences compared to the other two breeds, namely a higher concentration of polyunsaturated fatty acids, which can be related to being a fat-tailed breed. Even in restricted feeding conditions, this breed revealed the highest levels compared to Merino and Dorper sheep respectively, of linoleic acid (+31% and +28%), linolenic acid (+97% and +51%), eicosapentaenoic acid (EPA) (+65% and +37%), docosapentanenoic acid (DPA) (+31% Merino) and dodosahexanenoic acid (DHA) (+63% and +77%). EPA, DPA and DHA are three omega-3 fatty acids, with described beneficial characteristics. With this work we show other qualities (higher levels of the omega-3 fatty acids, EPA, DPA and DHA) of Damara meat that might present this breed as an interesting alternative for animal production in semi-arid climates.

Molecular characterisation of Mycoplasma species isolated from the genital tract of Dorper sheep in South Africa.

Biochemical and molecular analysis were conducted on 34 strains of Mycoplasma species isolated between 2003 and 2009 from the genital tract of clinically healthy Dorper sheep and sheep with ulcerative vulvitis and balanitis. Earlier publications identified the causative agent as Mycoplasma mycoides mycoides large colony (MmmLC) and Arcanobacterium pyogenes. The aims of the study were to characterise Mycoplasma species isolated from the genital tract of Dorper sheep with polymerase chain reaction assay, cloning and gene sequencing. Basic Local Alignment Search Tool (BLAST) results revealed six predominant Mycoplasma species: Mycoplasma arginini, Mycoplasma bovigenitalium, Arcanobacterium laidlawii, MmmLC, Mycoplasma sp. ovine/caprine serogroup II and M. canadense. Sequencing of the 34 isolates were analysed using phylogenetic methods, and 18 (50%) were identified as M. arginini with 99% – 100% similarity to M. arginini from England and Sweden. Six isolates showed 99% similarity to M. bovigenitalium strains from Turkey and Germany. Two isolates had 99% similarity to an M. sp. ovine/caprine sero group II from the United Kingdom. BLAST for two isolates revealed 99% similarity to Acholeplasma laidlawii from India, another two were 99% similar to MmmLC strain from Sweden, two showed 98% similarity to Mycoplasma sp. Usp 120 from Brazil, and two isolates have a 97% – 99% similarity to M. mm. Jcv1 strain from the United States of America. Finally, one isolate showed similarity of 99% to Mycoplasma canadense strain from Italy. The findings support the hypothesis that ulcerative vulvitis and balanitis of Dorper sheep in South Africa (SA) is a multifactorial disease with involvement of different Mycoplasma species.

Report of outbreaks of classical scrapie in Dorper sheep and associated prion protein gene polymorphisms in affected flocks.

Scrapie is an infectious neurodegenerative disease affecting sheep and goats, related with conformational alteration of an isoform of the prion protein that leads to deposition and aggregation in the host's central nervous system. Occurrence of the natural disease can be influenced by host genetic factors, such as a single nucleotide polymorphism of the prion protein gene. This study reports three scrapie-affected Dorper flocks located on three different farms in Brazil. The objective of this study was to analyze these three flocks using scrapie diagnostics, combining histology, immunohistochemistry, genotyping, and western blot assays. For immunohistochemistry, 192 sheep were selected and 308 sheep blood samples were taken for genotyping. A total of 22 sheep were scrapie positive by immunohistochemistry. Of these, four presented clinical signs and had scrapie immunoreactivity at the obex in western blot assays. The sheep without clinical signs were positive in lymphoid organs, such as the third eyelid and rectal mucosa. The major genotypes found on the flocks were ARQ/ARQ, ARQ/ARR, and ARQ/VRQ for codons 136, 154, and 171. Most of the sheep were considered to be at moderate to high risk, based on risk groups for developing scrapie. Some blood samples were sequenced, and polymorphisms were identified in other codons, such as 127, 142, and 143. Our data demonstrate the importance of preclinical scrapie diagnosis in Brazilian sheep, as most of the affected sheep showed no clinical signs, and emphasize the relevance of genotyping other Dorper sheep to determine the genotypic profile of the breed.

De novo assembly and characterization of the skeletal muscle transcriptome of sheep using Illumina paired-end sequencing

In order to enrich the ovine genome and provide a basis for future molecular genetics and functional genomics analyses in sheep, we used de novo assembly to establish transcriptomes of skeletal muscle tissues of Dorper and Small-tailed Han sheep. A total of 103,058,824 clean Illumina paired-end sequencing reads from the two libraries were assembled into 145,524 unigenes in a de novo project. There were 5718 unigenes showing differential expression between the two transcriptomes, and 7437 coding SSRs were exploited. After further assembly, we identified a total of 70,348 all-unigenes with an average length of 863bp; 35,201 of these all-unigenes could be annotated in the Nr database, and 12,219 were found in the clusters of orthologous groups database. Gene ontology searches indicated cell and binding as the main terms. Among 258 Kyoto Encyclopedia of Genes and Genomes database pathways, protein and amino acid metabolism pathways were the most commonly identified. We analyzed the ovine muscle transcriptome using high-throughput sequencing technology. Many unigenes were assembled and numerous molecular markers and differential expressed unigenes were identified.

Molecular cloning, characterisation and mRNA expression analysis of the sheep myosin light chain 1 gene

The complete cDNA sequence of the sheep MYL1 (Myosin light chain 1) gene was cloned using RT-PCR, 5′ RACE and 3′ RACE. We obtained two alternatively spliced isoforms of the MYL1 gene, MYL1a and MYL1b, which are 849 and 1046bp in length and encode proteins composed of 150 and 192 amino acid residues, respectively. And the GenBank accession numbers of MYL1a and MYL1b full-length cDNA sequences that we cloned are KJ700419 and KJ710701, respectively. Neither protein was predicted to have a signal peptide, but both were predicted to have several N-glycosylation and phosphorylation sites. More than half of the secondary structure of these proteins was predicted to be α-helical. The human MYL2 protein (1m8q.1.C) is the most similar in tertiary structure. Sequence alignment showed that the sheep MYL1a protein shares more than 92% amino acid sequence similar with Mus musculus, Homo sapiens, Rattus norvegicus, Sus scrofa and Gallus gallus and that the MYL1b protein shares more than 93% amino acid sequence similar with M. musculus, H. sapiens, R. norvegicus, Bos taurus and Oryctolagus cuniculus. Transcription profile analyses of various tissues indicated that the sheep MYL1a and MYL1b mRNAs were highly but differentially expressed in the longissimus dorsi. Moreover, the expression levels of these genes in the longissimus dorsi differed between Dorper and Small-tailed Han sheep. These results serve as a foundation for further investigations of the function of the sheep MYL1 gene.