绵羊regakine-1基因的克隆与序列分析

【研究目的】克隆并分析绵羊regakine-1基因;【方法】肠系膜淋巴结组织提取总RNA,利用设计的引物进行RT-PCR,PCR产物与pMD-19T载体连接后转化JM109感受态细胞,筛选阳性克隆、测序,并进行序列分析;【结果】克隆的绵羊regakine-1基因与牛的同源性为91%,推测的氨基酸序列信号肽为1￣21aa,SCY结构域为29￣87aa,结构特征与牛的相一致。【结论】克隆了绵羊regakine-1基因的ORF,并注册GenBank(AccessionEF617337)。

Molecular Cloning and Sequence Analysis of Ovine Regakine-1

OBJECTIVE]To clone and analysis the ovine regakine-1.METHOD]Total RNA was extracted from mesenteric lymph node and mRNA sequence of gene were amplified by RT-PCR with designed primers. The PCR products were ligated into the pMD-19T vector, and then transformed into competent cells of JM109. The recombinant plasmid was identified and the sequence was analyzed.RESULTS]Identity analysis showed that the regakine-1 nucleotide sequence shared 91% homology with that of cattle. The signal peptide of predicted amino acid sequence is 1~21aa and the structural domain SCY is 29~87aa, revealing that the structural feature is consistent with cattle regakine-1.CONCHUSION]The ovine ORF of regakine-1 gene was successfully cloned in present study and the sequence has been submitted to GenBank(Accession. EF617337).