AGPase基因植物过表达载体的构建及对甘蓝型油菜的转化

ADP-葡萄糖焦磷酸化酶（AGPase）是淀粉合成途径的关键酶。本研究克隆了大肠杆菌AGPase基因glg C,并通过双定点突变降低其对无机磷酸盐敏感性所致抑制作用以提高其酶活。而后将甘蓝型油菜AGPase导肽编码序列与glgc融合,构建成植物表达载体p MB-TG,并导入根癌农杆菌LB4404。利用农杆菌介导法转化甘蓝型油菜,获得转基因植株。PCR及半定量RT-PCR检测结果显示,该融合基因已导入油菜再生植株基因组并获得表达。增加淀粉积累为油脂合成提供更多碳源,被认为是一条潜在的提高油菜种子含油量的有效途径。本研究为利用遗传改良技术实现上述途径在油菜育种中的应用奠定前期基础。...

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Published in西南农业学报 Vol. 28; no. 3; pp. 975 - 981
Main Author 赵凯琴 罗延青 王敬乔 俎峰 陈苇 李劲峰
Format Journal Article
LanguageChinese
Published 云南省农业科学院经济作物研究所,云南昆明,650205 2015
Subjects
AGPase基因
植物表达载体
甘蓝型油菜
Brassica napus
AGPase gene
甘蓝型油菜
Plant expression vector
AGPase基因
植物表达载体
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ISSN1001-4829
DOI10.16213/j.cnki.scjas.2015.03.010

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Summary:ADP-葡萄糖焦磷酸化酶（AGPase）是淀粉合成途径的关键酶。本研究克隆了大肠杆菌AGPase基因glg C,并通过双定点突变降低其对无机磷酸盐敏感性所致抑制作用以提高其酶活。而后将甘蓝型油菜AGPase导肽编码序列与glgc融合,构建成植物表达载体p MB-TG,并导入根癌农杆菌LB4404。利用农杆菌介导法转化甘蓝型油菜,获得转基因植株。PCR及半定量RT-PCR检测结果显示,该融合基因已导入油菜再生植株基因组并获得表达。增加淀粉积累为油脂合成提供更多碳源,被认为是一条潜在的提高油菜种子含油量的有效途径。本研究为利用遗传改良技术实现上述途径在油菜育种中的应用奠定前期基础。
Bibliography:ZHAO Kai-qin, LUO Yan-qing, WANG Jing-qiao, ZU Feng, CHEN Wei, LI Jin-feng （Economic Crops Research Institute of Yunnan Academy of Agricultural Sciences, Yunnan Kunming 6S0205, China）
51-1213/S
ADP-giucese pyrophesphorylase（AGPase） is a key enzyme in starch synthesis. In this study, the gig（： of AGPsse gene from E. Coli was cloned, and enzymes activity was promoted by the inhibited effect caused by decreasing the sensitivity of inorganic phosphate through double site-directed mutations. The coding sequence d transit peptide from brassica napus was fused with gigc gene to construct a plant expression vector named pMB-TG, which was introduced into Agrobacterium tumefaciens LBA4404 and expressed itself into brassica napus plants. The results indicated that the construction of the AGPase gene plant expression vector was successful and nsehtl proved by mol- eculer detection. To increase the rapeseed starch accumulation for providing more carbon sources for the fat synthesis is an effective way to improve the oil con
ISSN:1001-4829
DOI:10.16213/j.cnki.scjas.2015.03.010