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当前位置: 首页 > 文章 > 干旱诱导基因GmNF-YA7克隆及植物表达载体构建 大豆科学 2022 (1) 43-48
Position: Home > Articles > Cloning and Plant Expression Vector Construction of Drought-Induced GmNF-YA7 Soybean Science 2022 (1) 43-48

干旱诱导基因GmNF-YA7克隆及植物表达载体构建

作  者：
殷书欣;计俊杰;肖情;李铭杨;何佳琦;张军;翟莹
单  位：
齐齐哈尔大学生命科学与农林学院/抗性基因工程与寒地生物多样性保护黑龙江省重点实验室;黑龙江省农业科学院畜牧兽医分院
关键词：
大豆;GmNF-YA7;干旱响应;植物表达载体;转基因工程菌
摘  要：
为了鉴定大豆核因子YA(Nuclear Factor YA, NF-YA)与非生物胁迫的关系,本研究检测大豆干旱胁迫下GmNF-YA7和GmNF-YA8的表达量情况,克隆GmNF-YA7基因并构建植物表达载体,同时获得其转基因工程菌株。qRT-PCR结果显示,GmNF-YA7和GmNF-YA8均可以被干旱胁迫诱导表达,且GmNF-YA7对干旱胁迫的应答更明显。从大豆中克隆出GmNF-YA7基因,其位于大豆8号染色体上,编码含有336个氨基酸的蛋白质,预测分子量为37.06 kDa, pI6.11。GmNF-YA7蛋白氨基酸序列中含有1个CBF保守结构域。亚细胞定位预测结果显示,GmNF-YA7定位于细胞核中。蛋白系统进化分析表明GmNF-YA7蛋白与拟南芥AtNF-YA1蛋白的亲缘关系较近。利用限制性内切酶Nde I和Sal I将GmNF-YA7与植物表达载体pRI101连接并转化农杆菌EHA105,获得转基因工程菌株。
译  名：
Cloning and Plant Expression Vector Construction of Drought-Induced GmNF-YA7
作  者：
YIN Shu-xin;JI Jun-jie;XIAO Qing;LI Ming-yang;HE Jia-qi;ZHANG Jun;ZHAI Ying;College of Life Science and Agro-Forestry of Qiqihar University/Heilongjiang Provincial Key Laboratory of Resistance Gene Engineering and Biodiversity Protection in Cold Regions;Branch of Animal Husbandry and Veterinary, Heilongjiang Academy of Agricultural Sciences;
关键词：
soybean;;GmNF-YA7;;drought response;;plant expression vector;;genetically engineered strain
摘  要：
In order to identify the relationship between soybean nuclear factor YA(NF-YA) and abiotic stress, we detected the expression levels of GmNF-YA7 and GmNF-YA8 in soybean under drought stress, cloned the GmNF-YA7, constructed the plant expression vector, and obtained genetically engineered strain in this reaserch. qRT-PCR results showed that both expression levels of GmNF-YA7 and GmNF-YA8 could be induced by drought stress, and the response of GmNF-YA7 to drought stress was more obvious. GmNF-YA7 was cloned from soybean, it was located on chromosome 8 of soybean and encoded a protein containing 336 amino acids. The predicted molecular weight of GmNF-YA7 protein was 37.06 kDa and the pI was 6.11. The amino acid sequence of GmNF-YA7 protein contained a conserved CBF domain. Subcellular localization prediction results showed that GmNF-YA7 protein was localized in the nucleus. Phylogenetic analysis showed that GmNF-YA7 protein was closely related to Arabidopsis AtNF-YA1 protein. GmNF-YA7 was constructed into plant expression vector pRI101 with restriction endonucleases Nde I and Sal I and transformed into Agrobacterium EHA105. This study showed that the GmNF-YA7 gene was up regulated by drought, and we constructed plant expression vector with GmNF-YA7 and genetically obtained engineered Agrobacterium strain with this vector.

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