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 | |
| Format | Journal Article |
| Language | Chinese |
| Published |
云南省农业科学院经济作物研究所,云南昆明,650205
2015
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| Subjects | |
| Online Access | Get full text |
| ISSN | 1001-4829 |
| DOI | 10.16213/j.cnki.scjas.2015.03.010 |
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| Abstract | 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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| AbstractList | S565.4; 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“卵,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pMB-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. 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。 |
| Author | 赵凯琴 罗延青 王敬乔 俎峰 陈苇 李劲峰 |
| AuthorAffiliation | 云南省农业科学院经济作物研究所,云南昆明650205 |
| AuthorAffiliation_xml | – name: 云南省农业科学院经济作物研究所,云南昆明,650205 |
| Author_FL | CHEN Wei LI Jin-feng ZU Feng WANG Jing-qiao ZHAO Kai-qin LUO Yan-qing |
| Author_FL_xml | – sequence: 1 fullname: ZHAO Kai-qin – sequence: 2 fullname: LUO Yan-qing – sequence: 3 fullname: WANG Jing-qiao – sequence: 4 fullname: ZU Feng – sequence: 5 fullname: CHEN Wei – sequence: 6 fullname: LI Jin-feng |
| Author_xml | – sequence: 1 fullname: 赵凯琴 罗延青 王敬乔 俎峰 陈苇 李劲峰 |
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| ClassificationCodes | S565.4 |
| ContentType | Journal Article |
| Copyright | Copyright © Wanfang Data Co. Ltd. All Rights Reserved. |
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| DOI | 10.16213/j.cnki.scjas.2015.03.010 |
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| DocumentTitleAlternate | Expression Vector Construction of AGPase and Its Utilization in Rapeseed（Brassica napus L. ） |
| DocumentTitle_FL | Expression Vector Construction of AGPase and Its Utilization in Rapeseed(Brassica napus L.) |
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| GrantInformation_xml | – fundername: 十二五“863”计划课题“强优势油菜杂交种的创制与应用”; 云南省自然科学基金面上项目 funderid: (2011AA10A104); (2010ZC171) |
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| Keywords | Brassica napus AGPase gene 甘蓝型油菜 Plant expression vector AGPase基因 植物表达载体 |
| Language | Chinese |
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| Notes | 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 |
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| PublicationTitle | 西南农业学报 |
| PublicationTitleAlternate | Southwest China Journal of Agricultural Sciences |
| PublicationTitle_FL | Southwest China Journal of Agricultural Sciences |
| PublicationYear | 2015 |
| Publisher | 云南省农业科学院经济作物研究所,云南昆明,650205 |
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| Snippet | 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... S565.4; 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“卵,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.... |
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| SubjectTerms | AGPase基因 植物表达载体 甘蓝型油菜 |
| Title | AGPase基因植物过表达载体的构建及对甘蓝型油菜的转化 |
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