绿豆防御素基因的克隆、序列分析和植物表达载体的构建

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.

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

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.