绵羊intelectin基因cDNA克隆与序列分析

【研究目的】克隆并分析绵羊intelectin基因;【方法】十二指肠粘膜组织提取总RNA,分别以上游电子克隆的拼接体和下游保守区为模板设计引物进行RT-PCR,PCR产物与pMD-19T载体连接后转化E.coli JM109,筛选阳性克隆并测序;【结果】克隆的绵羊intelectin基因与牛、猪、人、大鼠的同源性分别为93%、87%、83%、79%,预测的氨基酸序列包含FBG结构域。【结论】成功克隆了绵羊intelectin-2基因,并注册GenBank(Accession.EF624459)。

Molecular Cloning and Sequence Analysis of Sheep Intelectin cDNA

Objective]To clone and analysis the sheep intelectin.Method]Total RNA was extracted from duodenal mucosa tissue and mRNA sequence of gene were amplified by RT-PCR using designed primers,the upstream of which was designed from the contig by silico cloning,the downstream of which was designed in conservative region.The PCR products were ligated into the pMD-19T vector,and then transformed into competent cells of E.coli JM109.The sequence was analyzed to identify the recombinant plasmid.Results]Identity analysis showed that the intelectin nucleotide sequence shared 93.3%,87.1%,82.9% and 78.6% homology with that of bovine(predicted),porcine,human and rat.The predicted peptide contained a FBG motif.Conclusion] The sheep intelectin-2 was successfully cloned and the sequence has been submitted to GenBank(Accession.EF624459).