Abstract
An investigation of differences in gene expression in the longissimus muscle of Meishan and Large White pigs was undertaken, using the mRNA display technique. A fragment of one differentially expressed gene was isolated and sequenced, whereupon the complete cDNA sequence was then obtained by using the rapid amplification of cDNA ends (RACE). The nucleotide sequence of the gene is not related to any known porcine gene. Sequence analysis revealed that the open reading frame of this gene encodes a protein with 322 amino acids, thus displaying high sequence identity with the PDZ binding kinase (PBK) of eleven other animal species – dog, horse, cattle, human, chimpanzee, crab-eating macaque, rhesus monkey, rat, mouse, gray short-tailed opossum and platypus, so it can be defined as the porcine PBK gene. This gene was finally assigned GeneID:100141310. Phylogenetic tree analysis revealed that the swine PBK gene has a closer genetic relationship with the PBK gene of platypus. Gene expression analysis of eight tissues of a Meishan x Large White cross showed that the porcine PBK gene is differentially expressed in various tissues. Our experiment established the primary foundation for further research on this gene.
Highlights
The mRNA differential display, first described by Liang and Pardee (1992), is a quick and efficient method for identifying and characterizing altered gene expression in different cell types
The differentially expressed gene band was recovered from the gel and used as the template for re-amplification, which was performed with the corresponding oligo primer and the arbitrary primers used in mRNA differential display
Tissue expression profile Semi-quantitative RT-PCR analysis of tissue expression profiles was carried out using tissue cDNAs of one adult Meishan x Large White pig-cross as the templates
Summary
The mRNA differential display, first described by Liang and Pardee (1992), is a quick and efficient method for identifying and characterizing altered gene expression in different cell types. Chinese indigenous pig breeds, such as Meishan, Erhualian and Tongcheng, often possess valuable traits, such as resistance to diseases, high fertility, good maternal qualities, unique product attributes, longevity and the ability to adapt to harsh conditions. European pig breeds, such as the Large White, Landrace and Duroc, are noted for high growth-rate and lean meat content (Pan et al, 2003). The detection of differences in gene expression between Chinese indigenous and European breeds, or finding the differentially expressed genes between the two types, may serve as a basis for understanding the molecular mechanisms of these phenotypic differences
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