珙桐rbcL基因的克隆与序列分析  

Cloning and Sequence Analysis of rbcL Gene from Davidia Involucrata

作　　者：季红春[1,2] 胡进耀[2] 姜立春[2] 邹利娟[2] 贺静[2] 苏智先[2] 

机构地区：[1]西华师范大学环境科学与生物多样性重点实验室,四川南充637002 [2]绵阳师范学院,四川绵阳621000

出　　处：《绵阳师范学院学报》2009年第8期54-59,共6页Journal of Mianyang Teachers＇ College

基　　金：国家自然科学基金项目(30670209)

摘　　要：根据已知植物rbcL基因设计引物,以珙桐叶片总DNA为模板,PCR扩增目的DNA片段,克隆入pDM19-T载体。阳性克隆鉴定后进行测序,序列分析结果表明:该基因片断长为1266 bp,包括1031 bp的编码区序列,编码343个氨基酸;其编码区氨基酸与烟草、菠菜、玉米、甘薯、拟南芥、水稻、葡萄、地钱和葡萄柚等9个物种的同源性94.13%以上,并构建了它们间的亲缘关系树。该基因片断的预测晶体结构与菠菜的最相近,推测Lys86、Lys60、Lys179等残基对酶的活性影响较大。Primer was designed according to the sequences of the rbcL genes which were logged in Gene- Bank, Total DNA was extracted by modified CTAB from mature leaves of Davidia Involucrata. RbcL gene was obtained by polymerase chain reaction and cloned into pDM19 -T vector. The positive clone identified by PCR was sequenced. The nucleotide sequence was 1266 bp long, including an ORF of 1031bp, encoding a putative 343 amino acids. The deduced amino acid sequence had a high identity above 94.13% with that of tobacco, spinach, sweet potato, corn, Arabidopsis thaliana, Oryza sativa, Vitis vinifera, Marchantia polymorpha and Citrus paradise. Thus, the molecular phylogenetic tree was established. The crystal structure of the rbcL indicated Lys86, Lys60 and Lys179 were related to the activity of Rubisco.

关 键 词：珙桐 RBCL基因 克隆 序列分析 

分 类 号：Q7[生物学—分子生物学]