珍稀濒危鸟类褐马鸡mtDNA的分子克隆及序列分析  

Cloning and Sequencing of mtDNA Control Region in Rare and Endangered Borwn-eared Pheasant Crossoptilon mantchuricum from Shanxi

作　　者：武玉珍[1] 赵春贵[2] 王孟本[1] 张峰[1] 

机构地区：[1]山西大学黄土高原研究所,山西太原030006 [2]化学生物学与分子工程教育部重点实验室,山西太原030006

出　　处：《山西大学学报（自然科学版）》2008年第1期104-107,共4页Journal of Shanxi University(Natural Science Edition)

基　　金：国家自然科学基金(30570284);山西省自然科学基金(2006011077)

摘　　要：利用改进的SDS裂解、氯仿∶异戊醇抽提的方法提取了褐马鸡肌肉基因组DNA,通过PCR扩增的方法特异性扩增了褐马鸡线粒体(mtDNA)控制区(D-loop)全序列片段,PCR产物经凝胶回收纯化后与载体pGEM-TEasy相连,然后转化感受态细胞DH-5α,在含X-gal、IPTG的氨苄LB平板上筛选阳性克隆,重组质粒经PCR和琼脂糖凝胶电泳检测后,对目的片段进行测序.结果表明,经PCR特异性扩增出的褐马鸡线粒体控制区全序列碱基数为1 237 bp,用Blast程序进行检索比较表明此扩增序列与GeneBank中发表的褐马鸡线粒体控制区序列同源性高达99%,且具有鸟类线粒体的标志性序列.The genomic DNA of muscle of Crossoptilon mantchuricum （brown--eared pheasant） was extracted by modified SDS digesting and extracting by hydroxybenzene, then the mitochondrial DNA （mtDNA） control region of C. rnantchuricurn fragment was amplified by transcription--PCR, after gel purification,PCR products were inserted into the pGEM--T Easy cloning vectors. Recombinant pGEM--T the mitochondrial DNA control region of C. rnantchuricurn fragment vector was transuded into competent cell DH--5. Clones containing the DNA fragment of interest were screened on the LB agar plate （containing Amp） with X--gal and IPTG. PCR were performed to identify the clone containing the mtDNA control region of C. rnantchuricurn fragment of interest, and then the mtDNA fragment of interest was sequenced. The results showed that the obtained 1 237 bp DNA fragment shared a homology of 90% with the mtDNA control region of C. rnantchuricurn sequence registered in GenBank, and had the symbol sequences of avian.

关 键 词：褐马鸡 线粒体控制区 克隆 

分 类 号：Q785[生物学—分子生物学]