绿豆防御素基因的克隆、序列分析和植物表达载体的构建  

Cloning and Sequencing of Vigna Radiation Defensin Gene and Construction of Plant Expression Vector

作　　者：缪建锟[1] 孙黎[1] 彭晓明[1] 祝建波[2] 

机构地区：[1]石河子大学生命科学学院,新疆石河子832003 [2]新疆兵团绿洲生态农业重点实验室,新疆石河子832003

出　　处：《石河子大学学报（自然科学版）》2006年第1期112-115,共4页Journal of Shihezi University(Natural Science)

摘　　要：从绿豆叶片提取总DNA中,通过PCR方法扩增出362bp的具有多种抗病、抗虫特性的绿豆防御素基因,并将其克隆到pGM-T easy vector,酶切图谱及DNA测序分析表明克隆的片段包含了完整的绿豆防御素基因的编码序列,与原序列同源性达到99.5%,蛋白质同源性达到100%。此基因编码的多肽由73个氨基酸组成,含有28个氨基酸的信号肽和8个半胱氨酸,可形成4个二硫键。我们用此基因构建了高效植物表达载体pBin438-LD。Vigna Radiation Defensin is a kind of protein which bears the function of anti-fungi antibacteria and anti-pest. The Vigna Radiation defensin gene was amplified from Vigna Radiation total DNA by PCR, and was cloned into pGM-T easy vector. DNA sequence analysis indicated that the cloned 362bp DNA fragment carries the entire defensin coding sequence,which encoded 73 amino acids, including 23 amino acids signal peptides, 8 cysteine which can form 4 pairs disulfide bonds. We cloned the Vigna Radiation Defensin gene which has the highest homology compared with the published data,showing 99.5% and 100% homo-logy at the nucleotide and at the amino acid sequence respectively. A Vigna Radiation Defensin gene plant expression vector has been constructed.

关 键 词：防御素 绿豆 植物表达载体 

分 类 号：Q785[生物学—分子生物学]