黄花苜蓿MfNHX基因和MfP5 CS基因的克隆和序列分析  

Cloning and Sequence Analysis of MfNHX and MfP5 CS Genes in Medicago falcata L.

作　　者：曾光[1] 石庆华[1] 张博[2] 许磊[1] 陈全家[1] 张桦[1] 

机构地区：[1]新疆农业大学农学院/农业生物技术重点实验室,乌鲁木齐830052 [2]新疆农业大学草业工程与环境科学学院/新疆草地资源与生态重点实验室,乌鲁木齐830052

出　　处：《新疆农业科学》2010年第6期1231-1235,共5页Xinjiang Agricultural Sciences

基　　金：国家转基因重大专项(2008ZX08005-004);国家科技支撑计划(2008BADB3B06)

摘　　要：【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。[ Objective]To obtain MfNHX and MfP5 CS 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 DH5a, 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 MfP5 CS is 97.63 %. Compared with Medicago. varia. Xinmu No. 1 on NCBI, the MfNHX homology of nueleotide sequence is 99.45%, and the MfP5 CS is 98.84%. [ Conclusion] The MfNHX and MfP5 CS genes have been obtained, which lays the foundation for further research on stress tolerance in alfalfa.

关 键 词：黄花苜蓿 MfNHX MfP5CS 克隆 序列分析 

分 类 号：S551.7[农业科学—作物学] S188