猪Musclin基因片段克隆与序列分析

【研究目的】克隆并分析猪musclin基因；【方法】肌肉组织提取总RNA，利用设计的引物进行RT-PCR，PCR产物与pMD19-T连接后转化E. coli DH5α，检测阳性克隆并测序；【结果】克隆的猪musclin基因片段与人、大鼠、小鼠同源性分别为86%、78%、75%，预测的氨基酸序列含有“KKKR”结构和与小鼠ANP、BNP、CNP蛋白的同源性区域；【结论】克隆了猪musclin基因片段并注册GenBank (Accession.EF369511)。

Molecular cloning and sequence analysis of porcine musclin

【Objective】To clone and analysis the porcine musclin.【Method】Total RNA was extracted from muscle tissue and mRNA sequence of gene were amplified by RT-PCR using two designed primers. The PCR products were ligated into the pMD-19Tvector, and then transformed into competent cells of E. coli DH5α.The sequence was analyzed to identify the recombinant plasmid.【Results】Identity analysis showed that the musclin nucleotide sequence shared 86%, 78% and 75% homology with that of human , rat and mouse. The predicted peptide contained a “KKKR” motif and the region homologous to mouse ANP, BNP, and CNP.【Conclusion】The porcine partial musclin gene was successfully cloned in present study and the sequence has been submitted to GenBank(Accession.EF369511).