RACE Technique Research Articles

‘Popping the Clutch’: Novel Mechanisms Regulating Sexual Development in Cryptococcus neoformans

Sexual reproduction in fungal pathogens such as Cryptococcus provides natural selection and adaptation of the organisms to environmental conditions by allowing beneficial mutations to spread. However, successful mating in these fungi requires a time-critical induction of signaling pheromones when appropriate partners become available. Recently, it has been shown that the fungus uses the transcriptional equivalent of the racing technique: 'popping the clutch'-pushing in the clutch pedal, putting the car in gear, revving with the gas pedal, and then dropping the clutch pedal to accelerate rapidly. In the same way, Cryptococcus during vegetative growth constitutively matches a high rate of pheromone synthesis with a high rate of degradation to produce repressed levels of transcript. Then, when mating is required, the fungus drops the degradative machinery, resulting in a rapid induction of the pheromone. Pairing with this novel regulatory cycle is a host of mitogen-activated protein kinase cascades, cyclic AMP-dependent, and calcium-calcineurin signaling pathways that maintain these high rates of pheromone synthesis and prime downstream pathways for an effective mating response. The intersection of a number of virulence-associated traits with sexual development such as the synthesis of an immune-disruptive laccase as well as a protective polysaccharide capsule makes these rapid regulatory strategies a formidable foe in the battle against human disease.

Cloning of vacuolar H+-ATPase subunit c genes from Japanese iris, and functional characterization in yeast

Five different isoforms (IrlVHA-c1-c5) of V-ATPase subunit c (VHA-c) were cloned from a Japanese iris (Iris lactea Pall. var. chinensis Fisch. Koidz) cDNA library using degenerate primers PCR and the 5′-RACE technique. The sequence analysis showed the open reading frame (ORF) of the IrlVHA-c1-c5 to be 495 bp, corresponding to a protein of 164 amino acids. Among the five isoforms, IrlVHA-c1 and IrlVHA-c2 are completely homologous. The IrlVHA-c protein is localized at the vacuolar membrane as indicated by a green fluorescent protein (GFP) marker. Its over-expression in yeast could enhance yeast tolerance to NaCl stress. These results show that there are at least five genes encoding different isoforms of IrlVHA-c in Japanese iris and IrlVHA-c is important for the function of V-ATPase.

Molecular characterization of a cucumber nitrate reductase (CsNR) gene under NO3 − stress

Nitrate reductase is a key enzyme in the overall process of nitrate assimilation by plants. A full-length cDNA clone encoding nitrate reductase (NR; EC 1.6.6.1) was isolated from cucumber (Cucumis sativus L.) by RT-PCR and RACE techniques. The NR of cucumber (CsNR), a full-length cDNA sequence of 3032bp contains an open reading frame of 2748bp encoding 915 amino acids. The deduced 915 amino acid sequence showed high identities with NR from other plants. Quantitative real-time PCR analysis indicated that CsNR expression was different in root, stem, leaf, flower and mature fruit tissues. CsNR transcript level and nitrate reductase activity (NRA) was down-regulated and the change in NO(3) (-) concentration showed a negative trend with NRA in leaves when subjected to the 182mM NO(3) (-) treatment. However, the CsNR transcript level was up-regulated in roots by 182mM NO(3) (-) treatment. Furthermore, NRA in roots lagged behind CsNR expression and there was no obvious lag of NRA in leaves. This study found that in roots, there was no obvious relationship between NRA and NO(3) (-) content. These results indicated that NRA was not only controlled by the level of CsNR mRNA and there was an obvious negative relationship between NO(3) (-) content and NRA in leaves but not in roots.

Cloning and transcription analysis of the Candida rugosa propionyl‐CoA dehydrogenase gene and its expression in Pichia pastoris

The propionyl-CoA dehydrogenase (PACD) gene was cloned from Candida rugosa by the cDNA RACE technique. The full cDNA of the PACD gene has a length of 1408 bp, which contains a complete open reading frame (ORF) of 1329 bp, coding for 442 amino acids. The cDNA of PACD was cloned into the expression plasmid pPIC9K and transformed into Pichia pastoris GS115. The recombinant protein was purified by Ni-NTA affinity chromatography, and its size was observed to be approximately 49 kDa as estimated by SDS-PAGE. Anti-His antibodies were used to characterise the recombinant PACD by western-blot analysis. The recombinant protein retained the activity of catalysing propionyl-CoA to acryloyl-CoA. The results of dot-blotting hybridisation using a PACD cDNA probe indicated that the PACD mRNA level was modified at different stages: mRNA levels were low for the first 36 h, then increased through 48 h and eventually reached a stable level. These results indicate that propionate induction could significantly activate PACD mRNA expression. Information from this study will be helpful in elucidating the metabolic pathway for 3-hydroxypropionic acid production in C. rugosa.

Expression of mitochondrial malate dehydrogenase in Escherichia coli improves phosphate solubilization

Penicillium oxalicum C2 was isolated from the rhizosphere of Mexican stonecrop based on its high level of tricalcium phosphate solubilizing activity. A full-length gene encoding mitochondrial malate dehydrogenase (mMDH) was cloned from P. oxalicum C2 by RT-PCR and RACE techniques. The cDNA was 1,284 bp in length, with a complete open reading frame of 1,023 bp encoding a protein of 340 amino acids with a predicted molecular mass of 35.7 kDa. The predicted amino acid sequence shared high identity with mMDHs from other organisms. Escherichia coli strain BL21(DE3) transformed with the P. oxalicum C2 mMDH gene showed an increase in MDH activity of about 5-fold compared to non-transformed cell as measured in cell extracts. It also secreted more malate, lactate, acetate, citrate, oxalate and produced higher clearing halos when grown in plates with tricalcium phosphate as the sole source of phosphate. These results showed that expression of the P. oxalicum C2 mMDH gene in E. coli could enhance organic acid secretion and improve the phosphate solubilizing ability of the cell.

Cloning RPL11 cDNA 3′ End by SMART RACE Technique

RT-PCR and RACE techniques were used to clone 3′ end real sequence of RPL11 gene from a goat. The sequences included some non-structural protein (3D) genes, and 3D gene immediately downstream 3′ non-coding region (UTR) and poly (A) tail. The results showed that the length of specific amplified fragment was 566 bp (not including polyA tail). RPL11 gene encoded 188 amino acids, amino acid sequence was conducted for homology analysis by NCBI BLAST tool. The highest homology was Mus (RPL11, mRNA homology 99%), followed by Bos (RPL11, mRNA homology 96%), and the next was Hs (RPL11, mRNA homology of 93%).

Comprehensive Analysis of Transcript Start Sites in Ly49 Genes Reveals an Unexpected Relationship with Gene Function and a Lack Of Upstream Promoters

Comprehensive analysis of the transcription start sites of the Ly49 genes of C57BL/6 mice using the oligo-capping 5′-RACE technique revealed that the genes encoding the “missing self” inhibitory receptors, Ly49A, C, G, and I, were transcribed from multiple broad regions in exon 1, in the intron1/exon2 region, and upstream of exon -1b. Ly49E was also transcribed in this manner, and uniquely showed a transcriptional shift from exon1 to exon 2 when NK cells were activated in vitro with IL2. Remarkably, a large proportion of Ly49E transcripts was then initiated from downstream of the translational start codon. By contrast, the genes encoding Ly49B and Q in myeloid cells, the activating Ly49D and H receptors in NK cells, and Ly49F in activated T cells, were predominantly transcribed from a conserved site in a pyrimidine-rich region upstream of exon 1. An ∼200 bp fragment from upstream of the Ly49B start site displayed tissue-specific promoter activity in dendritic cell lines, but the corresponding upstream fragments from all other Ly49 genes lacked detectable tissue-specific promoter activity. In particular, none displayed any significant activity in a newly developed adult NK cell line that expressed multiple Ly49 receptors. Similarly, no promoter activity could be found in fragments upstream of intron1/exon2. Collectively, these findings reveal a previously unrecognized relationship between the pattern of transcription and the expression/function of Ly49 receptors, and indicate that transcription of the Ly49 genes expressed in lymphoid cells is achieved in a manner that does not require classical upstream promoters.

4-Coumarate:CoA ligase family members from elicitor-treated Sorbus aucuparia cell cultures

Sorbus aucuparia cell cultures accumulate biphenyl and dibenzofuran phytoalexins in response to elicitor treatment. These polyketide derivatives arise from the starter substrate benzoyl-CoA, the biosynthesis of which is largely unresolved. Two CoA ligases involved are cinnamate:CoA ligase and benzoate:CoA ligase, which were assumed to be related in S. aucuparia to the ubiquitous 4-coumarate:CoA ligase (4CL). cDNAs encoding three distinct 4CLs from elicitor-treated S. aucuparia cell cultures were isolated using RT-PCR and RACE techniques and functionally expressed in Escherichia coli as His 6-tagged proteins (Sa4CL2 and Sa4CL3) or GST-fusion protein (Sa4CL1). All three isoenzymes preferred 4-coumaric acid over cinnamic acid in spectrophotometric assays and failed to utilize benzoic acid in radioisotopic assays. After elicitor treatment of S. aucuparia cell cultures, the transcript levels of all three Sa4CLs increased but were significantly lower than the maximum expression rates of the phenylalanine ammonia-lyase ( PAL) and biphenyl synthase 1 ( BIS1) genes. The substrate specificities and the expression profiles indicate that the three 4CL isoenzymes are not involved in benzoyl-CoA biosynthesis in S. aucuparia cell cultures. Sa4CL3 and PAL transcripts also accumulated in response to light treatment. Phylogenetically, Sa4CL1 and Sa4CL2 belong to the class I cluster and Sa4CL3 groups in the class II cluster. Sa4CL3 contains a 49-amino acid N-terminal extension, which includes a chloroplast sorting signal.

Transcriptional regulation of two RTE-like genes of carnation during flower senescence and upon ethylene exposure, wounding treatment and sucrose supply

RTE1 (REVERSION-TO-ETHYLENE SENSITIVITY1) was identified as a positive regulator of ETR1 (ethylene resistant1) function in Arabidopsis; RTEs are a small gene family. Ethylene plays a crucial role in the senescence of carnation (Dianthus caryophyllus L.) flowers. Two cDNA clones encoding putative RTE-like protein (DCRTE1 and DCRTH1) were obtained from total RNA isolated from senescing carnation petals using RT-PCR and RACE techniques. The predicted proteins of DCRTE1 and DCRTH1 consist of 228 and 233 amino acids, respectively. Interestingly, the deduced DCRTE1 protein, like most other RTEs, includes two putative transmembrane domains, while the deduced DCRTH1 protein includes five putative transmembrane domains, according to the TMHMM database. Northern blots showed that the level of DCRTE1 mRNA in petals first decreased then increased remarkably after ethylene production started, and DCRTE1 expression showed an increasing trend in ovaries during natural flower senescence. The amount of DCRTH1 transcripts increased gradually in both petals and ovaries during natural senescence. Exogenous ethylene increased transcript abundance of DCRTE1 and DCRTH1 to various degrees in both petals and ovaries. STS treatment decreased the level of DCRTH1 mRNA in petals and ovaries compared with the control. DCRTE1 and DCRTH1 showed a rapid increase and then a decrease in mRNA accumulation in leaves after wounding. These results suggest that both DCRTE1 and DCRTH1 could play important roles in flower senescence-related signalling. Sucrose treatment did not remarkably affect the amount of DCRTE1 and DCRTH1 mRNAs.

Particle-in-cell simulations with charge-conserving current deposition on graphic processing units

We present an implementation of a 2D fully relativistic, electromagnetic particle-in-cell code, with charge-conserving current deposition, on parallel graphics processors (GPU) with CUDA. The GPU implementation achieved a one particle-step process time of 2.52ns for cold plasma runs and 9.15ns for extremely relativistic plasma runs, which are respectively 81 and 27 times faster than a single threaded state-of-art CPU code. A particle-based computation thread assignment was used in the current deposition scheme and write conflicts among the threads were resolved by a thread racing technique. A parallel particle sorting scheme was also developed and used. The implementation took advantage of fast on-chip shared memory, and can in principle be extended to 3D.

Cloning and characterization of Caveolin-1 gene in pigeon, Columba livia domestica

Caveolins, a class of principal proteins forming the structure of caveolae in plasmalemma, were encoded by caveolins gene family. Caveolin-1 gene is a member of caveolins gene family. In the present study, a full-length of 2605 bp caveolin-1 cDNA sequence in Columba livia domestica, which included a 537 bp complete ORF encoding a 178 amino acids long putative peptide, were obtained by using RT-PCR and RACE technique. The Columba livia domestica caveolin-1 CDS shared 80.1% - 93.4% homology with Bos taurus, Canis lupus familiaris, Gallus gallus and Rattus norvegicus. Meanwhile, the putative amino acid sequence of Columba livia domestica caveolin-1 shared 85.4% - 97.2% homology with the above species. The semi-quantity RT-PCR revealed that Caveolin-1 expressions were detectable in all the Columba livia domestica tissues and the expressional level of caveolin-1 gene was high in adipose, medium in various muscles, low in liver. These results demonstrated that Caveolin-1 gene was potentially involved in some metabolic pathways in adipose and muscle.

The impact of radical cystectomy on renal function: An analysis of variance based on age, gender, race and diversion technique

The impact of radical cystectomy on renal function: An analysis of variance based on age, gender, race and diversion technique

Relationship between Rh-RTH1 and ethylene receptor gene expression in response to ethylene in cut rose

A cDNA clone encoding a putative RTE1-like protein (Rh-RTH1) was obtained from total RNA isolated from senescing rose (Rosa hybrida cv. Tineke) petals using RT-PCR and RACE techniques. The cDNA (1,061 bp) contained an open reading frame of 684 bp corresponding to 227 amino acids. The amino acid sequence had 60.0, 49.6, 61.2, 42.5 and 39.8% identity with that of Arabidopsis RTH, RTE1, tomato GRL2, GRL1 and GR, respectively. Northern hybridization indicated that Rh-RTH1 expression is enhanced by endogenous and exogenous ethylene and inhibited by 1-MCP in petals and gynoecia. Rh-RTH1 expression partly correlated with sites of the ethylene receptor gene Rh-ETR1 and Rh-ETR3 expression, such as the petals, gynoecia, roots, and buds. The induction of Rh-RTH1 and Rh-ETR3 expression was substantially suppressed by 1-MCP treatment, while Rh-ETR1 expression was not reduced by 1-MCP treatment. Following treatment of flowers with sucrose, the level of Rh-RTH1 and Rh-ETR3 mRNA was only slightly decreased in petals and gynoecia. Upon wounding treatment, Rh-RTH1, Rh-ETR1 and Rh-ETR3 showed a quick increase in mRNA accumulation which was positively correlated with the increase in ethylene production. The expression of Rh-RTH1 showed partial correlation with that of Rh-ETR1 and Rh-ETR3.

Cloning and functional analysis of trehalose -6-phosphate synthase gene from <I>Selaginella pulvinata</I>

Trehalose-6-phosphate synthase, a key enzyme in trehalose synthesis pathway of plant, plays an important role in response to abiotic stress in xerophilous rock lily and other resurrection plants. In this study, homologous amplification and RACE technique were used to clone gene SpTPS1 for trehalose-6-phosphate synthase from Selaginella pulvinata, which is an endemic xerophilous plant in China. The full-length cDNA is 3223 bp long, containing an open reading frame (ORF) of 2790 bp. Protein sequence comparison showed that the pupative amino acid sequence of this ORF shares high similarity to trehalose-6-phosphate synthases of mode species, especially at the conserved sites of catalytic activity centers. Yeast functional complementation test showed that trehalose-6-phosphate synthase mutant (tps1 big up tri, open), transformed by the ORF of SpTPS1 gene, can restore growth on the medium supplemented with glucose as a sole carbon source. This result indicated that SpTPS1 of S. pulvinata encodes for an active protein and is hopeful to be applied in transgenic improvement of abiotic stress tolerance in plant.

Molecular cloning, characterization and mRNA expression of selenium-dependent glutathione peroxidase from abalone Haliotis discus hannai Ino in response to dietary selenium, zinc and iron

A novel selenium-dependent glutathione peroxidase (Se-GPX) was cloned from abalone Haliotis discus hannai Ino (HdhGPx) by homology cloning with degenerate primers and RACE techniques. The full length of HdhGPx cDNA was 963bp with a 669bp open reading frame (ORF) encoding 222 amino acids and a 101bp eukaryotic selenocysteine insertion sequence (SECIS) in 3′ untranslated region (UTR). It was showed that HdhGPx has a characteristic codon at 235TGA237 that corresponds to selenocysteine (SeC) as U72. Sequence characterization revealed that HdhGPx contains a characteristic GPx signature motif 2 (96LGLPCNQF103), an active site motif (179WNFEKF184). In addition, two potential N-glycosylation sites (112NGTE115 and 132NLTQ135) were identified in HdhGPx. 3D modeling analysis showed that the overall structure of HdhGPx monomer had more similarity to human GPx3 than human GPx1. Relatively higher-level mRNA expression was detected in hepatopancreas, mantle and gonad by real-time PCR assays. The relative expression levels of HdhGPx mRNA in hepatopancreas and haemocytes were detected by real-time PCR in abalone fed with nine different diets containing graded levels of selenium (0.15, 1.32 and 48.7mgkg−1), zinc (6.69, 33.85 and 710.63mgkg−1) and iron (29.17, 65.7 and 1267.2mgkg−1) for 20weeks, respectively. The results showed that the expressions of HdhGPx mRNA were statistically higher at adequate dietary selenium (1.32mgkg−1), zinc (33.85mgkg−1) and iron (65.7mgkg−1) than those in low dietary minerals, respectively. But HdhGPx mRNA expression levels were down-regulated by high contents of dietary selenium (48.7mgkg−1), zinc (710.63mgkg−1) and iron (1267.2mgkg−1), respectively. These results indicated that adequate dietary minerals could increase the mRNA expression of HdhGPx, and then to increase the total antioxidant capacities in abalone.

Transcriptional Regulation and Molecular Characterization of the manA Gene Encoding the Biofilm Dispersing Enzyme Mannan endo-1,4-β-Mannosidase in Xanthomonas campestris

Exopolysaccharide and several extracellular enzymes of Xanthomonas campestris pv. campestris (Xcc), the causative agent of black rot in crucifers, are important virulence determinants. It is known that Clp (cAMP receptor protein-like protein) and RpfF (an enoyl-CoA hydratase homologue required for the synthesis of diffusible signal factor, DSF) regulate the production of these determinants. Addition of DSF or Xcc extracellular protein containing partially purified mannanase (EC 3.2.1.78, encoded by manA) can disperse the cell aggregates formed by rpfF mutant. In this study, nucleotide G 64 nt upstream of the manA translation start codon was determined as the transcription initiation site by the 5' RACE technique. Transcriptional fusion assays showed that manA transcription is positively regulated by Clp and RpfF and induced by locust bean gum. The manA coding region was cloned and expressed in E. coli as recombinant ManA (rManA). The rManA was purified by affinity chromatography, and its biochemical properties were characterized. The rManA had a pH optimum at 7.0 (0.1 M Hepes) and a temperature optimum at about 37 degrees C. Sequence and mutational analyses demonstrated that Xcc manA encodes the major mannanase, a member of family 5 of glycosyl hydrolases. This study not only extends previous work on Clp and RpfF regulation by showing that they both influence the expression of manA in Xcc, but it also for the first time characterizes Xanthomonas mannanase at the protein level.

Canine CXCL7 and its functional expression in dendritic cells undergoing maturation

Many cells, including leucocytes and stromal cells, express CXCL7, a member of the CXC chemokine family, also known as platelet basic protein. CXCL7 is a potent chemoattractant and activator of neutrophil function. Dendritic cells (DCs) play a pivotal role in antigen processing and presentation. Very little information is available on the ability of DCs to recruit neutrophils by producing chemokines. In this work, we have cloned canine CXCL7. Based on the predicted gene sequence and using the 3′RACE technique, the full-length gene was amplified from LPS-treated canine peripheral blood mononuclear cells. The cloned cDNA sequence consisted of 357 nucleotides and encoded a 118 amino acid protein, including a 38 amino acid signal peptide. The use of CXCL7-containing supernatants from CXCL7-transfected BALB/3T3 in the neutrophil migration assay confirmed that canine CXCL7 had chemoattractive activity for neutrophils. We then used canine monocyte-derived DCs to generate CXCL7 for the rest of the experiment. Expression of CXCL7 by DCs treated with LPS, IL-1β, IL-6, TGF-β, TNF-α, or IFN-γ was compared using real-time RT-PCR and Western blotting. When treated with IL-1β, IL-6, TNF-α, or TGF-β, canine DCs expressed significantly higher levels of CXCL7 mRNA and protein than when treated with IFN-γ or LPS. It is concluded that dog DCs express high levels of the neutrophil chemotactic factor CXCL7 when stimulated by proinflammatory cytokines, including IL-1β, IL-6, TNF-α, or TGF-β, and may play an important role in modulating inflammatory responses.

Cloning and expression analysis of a ribosomal protein S7 in wheat under the stress of Puccinia triticina

A gene was isolated from the suppression subtractive hybridization (SSH) cDNA library of wheat by reverse northern blotting. Full length cDNA was obtained by RACE technique, and the result showed that all sequences consisted of 471 bp and encoded 157 amino acid. Subsequently, the gene expression was enhanced according to quantity real-time PCR (qRT-PCR) in response to leaf rust stress as the time over. All the results above suggested that the gene might play important role in wheat hypersensitive response (HR) to leaf rust.

Prognostic Value of Vascular Endothelial Growth Factor Expression in Patients with Lung Cancer: A Systematic Review with Meta-Analysis

Vascular endothelial growth factor (VEGF) has been implicated in tumorigenesis and metastasis, and it presumably mediates the proliferation of endothelial cells and promotes vascular permeability. However, the prognostic value of VEGF overexpression in patients with lung cancer remains controversial. Survival data from published studies were aggregated following a methodological assessment. A systematic review of eligible studies with meta-analysis was performed to quantitatively review the correlation of VEGF overexpression with survival in patients with lung cancer. We conducted a final analysis of 5386 patients from 51 studies. The studies were categorized by histology, disease stage, patient race, VEGF isoform, and laboratory techniques used. Combined hazard ratios suggested that VEGF overexpression had an unfavorable impact on survival of patients with non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC). However, VEGFC and vascular endothelial growth factor receptor 3 (VEGFR3)/flt-1 overexpression did not significantly correlate with survival in patients with NSCLC. In stage I-III NSCLC with VEGF, the hazard ratio (95% confidence interval) was 1.46 (1.38-1.54) overall, 1.35 (1.24-1.46) in Asian patients, 1.61 (1.49-1.73) in non-Asian patients, 1.41 (1.17-1.65) in SCLC, 1.27 (1.06-1.47) in adenocarcinoma, 1.57 (1.43-1.70) in stage I NSCLC, 1.46 (1.38-1.55) in NSCLC by immunohistochemistry, 1.52 (1.23-1.81) in NSCLC by reverse transcription-polymerase chain reaction, 1.22 (0.96-1.47) in NSCLC with VEGFC, and 1.58 (0.96-2.20) in NSCLC with VEGFR3/flt-1. The data collected were not sufficient to determine the prognostic value of VEGF in patients with squamous cell lung carcinomas. VEGF overexpression indicates a poor prognosis for patients with NSCLC and SCLC; VEGFC and VEGFR3/flt-1 overexpression was not significantly correlated with survival for patients with NSCLC.

Characterization and expression of bone morphogenetic protein 4 gene in postnatal pigs

Bone morphogenetic protein 4 (BMP4) is involved in animal embryonic development and reproductive physiology. The human and murine BMP4 genes have been isolated and characterized. The objectives of this study were to: (1) characterize the full mRNA and genomic sequence for porcine BMP4, and (2) examine BMP4 gene expression in 10 tissues of postnatal female pigs. Using RT-PCR, RACE and general PCR techniques, a 1,626 bp DNA including the full coding region of BMP4 was isolated and identified as a homologue of human BMP4 transcript variant (TV)-c. The porcine TV-c contained 3 exons and astride 3.6 kb in the isolated 7.8 kb porcine BMP4 genome. The In silicon cloning identified other three forms of mRNAs, including the homologues of human TV-1, TV-a and a novel variant related to human TV-3 (TV-3p). The porcine TV-c, TV-1 and TV-3p bear internal ribosome entry sites (IRES) in 5' untranslated region (UTR), while there are two ARE elements in the 3'UTR. The full genomic sequence of porcine BMP4 gene showed 81.38, 76.23 and 64.00% identity with that of bovine, human and murine, respectively. The expression of BMP4 mRNA was determined by RT-PCR in 7, 14, and 28 day old female piglets and non-gestational sows. The results showed that porcine BMP4 occurred in all 10 examined tissues (heart, lung, liver, kidney, ovary, spleen, spinal medulla, brain, duodenum and thymus). The mRNA expression levels were relatively higher in lung and kidney in 7 day old piglets, thymus in 14 day old piglets, and spleen in 28 day old piglets, respectively, while the higher expressions were detected in liver of non-gestational pigs (P < 0.05). Moreover, the mRNA amounts both in 7 day old piglets and sows were generally higher than those in 14 and 28 day old piglets in nearly all examined tissues, except in thymus. It is concluded that the structure of porcine BMP4 gene is highly conservative with other mammalian BMP4 genes, but some differences may present in the regulation of gene expression. BMP4 mRNAs are expressed in postnatal pigs, and is spatiotemporally regulated.