黄花苜蓿MfNHX基因和MfP5CS基因的克隆和序列分析

目的]获得黄花苜蓿(Medicago falcata L.)的MfNHX基因和MfP5CS基因,并对这两个基因进行序列分析.方法]利用Trizol总RNA提取试剂盒提取了新疆野生黄花苜蓿的总RNA,并用特异引物对其进行RT-PCR扩增,将得到的cDNA片段连接到pMD19-T载体上,最后将该载体转化大肠杆菌DH5α,并对阳性克隆进行了序列分析.结果]MfNHX基因全长1 626 bp,Genbank登录号为GU265772;MfP5CS基因全长2 148 bp,Genbank登录号为GU180149.它们与紫花苜蓿核苷酸序列的同源性分别为99.63;和97.63;,与新牧1号杂花苜蓿核苷酸序列的同源性分别为99.45;和98.84;.结论]研究已经获得MfNHX基因和MfP5CS基因,为进一步进行苜蓿抗逆境胁迫的研究奠定了基础.

Cloning and Sequence Analysis of MfNHX and MfP5CS Genes in Medicago falcata L.

【Objective】To obtain MfNHX and MfP5CS genes in Medicago falcata L.and analyze their sequences.【Method】The total RNA of wild Medicago falcata L.in Xinjiang was extracted with Trizol Total RNA Extracting Reagent Kit and amplified to cDNA by RT-PCR using special primer.The pMD19-T vector linked with the cDNA fragment was transformed into Escherichia coli DH5α,and the positive clone was sequenced.【Result】The MfNHX(GenBank accession number: GU265772) is 1 626 bp,the MfP5CS(GenBank accession number: GU180149) is 2 148 bp.Compared with Medicago.sativa on NCBI,the MfNHX homology of nucleotide sequence is 99.63%,and the MfP5CS is 97.63%.Compared with Medicago.varia.Xinmu No.1 on NCBI,the MfNHX homology of nucleotide sequence is 99.45%,and the MfP5CS is 98.84%.【Conclusion】The MfNHX and MfP5CS genes have been obtained,which lays the foundation for further research on stress tolerance in alfalfa.