蛋氨酸裂解酶的基因克隆及序列分析  

Cloning of L-Methionine γ-lyase gene

作　　者：马百坤[1] 唐晓莹[2] 黄惠臣 时永辉[1] 

机构地区：[1]南京军区南京总医院解放军检验医学研究所,江苏南京210002 [2]昆山市第三人民医院检验科,江苏昆山215316 [3]张家港市第一人民医院检验科,江苏张家港215611

出　　处：《医学研究生学报》2006年第12期1063-1066,共4页Journal of Medical Postgraduates

基　　金：江苏省医学重点学科基金资助项目(苏卫科教[2001]34)

摘　　要：目的:克隆蛋氨酸裂解酶(METase)中阴道毛滴虫蛋氨酸裂解酶(TVMGL1)基因,探讨重组METase(rMETase)对肿瘤细胞生长的抑制作用。方法:从培养的阴道毛滴虫提取总RNA,以此为模板,利用RT-PCR获取TVMGL1基因,琼脂糖电泳分离纯化所得目的基因并克隆至T-Vector,AB I 3730自动序列分析仪分析所得基因序列。结果:PCR产物经琼脂糖电泳,目的基因片段大小与预期大小相符;序列分析所得基因序列为1 191 bp,与国外报导的序列相符。结论:从培养的阴道毛滴虫中成功克隆出TVMGLV1基因,为METase抗肿瘤作用的研究奠定了基础。Objective：To identify, clone, sequence the L-Methionine γ-lyase gene from Trichomonas vaginalis（ TVMGL1 ）. Methods：Total RNA was prepared from primitive protozoon trichomonas vaginalis, cDNA fragment encoding Methionine γ-lyase gene was amplified by RT-PCR using specific primers, and then was cloned into pGEM-T vector. Inserted Methionine γ-lyase gene was sequenced by ABI 3730 DNA Sequencer. Results：Analysis indicates that the DNA fragment is 1 191 base pairs in length and has high nucleotide homology with that reported previously. Conclusion：L-Methionine γ-lyase gene was successfully cloned.

关 键 词：蛋氨酸裂解酶 基因克隆 反转录PCR 

分 类 号：Q785[生物学—分子生物学]