gpd1基因莱茵衣藻表达载体构建及农杆菌介导转化莱茵衣藻  

Construction of gpd1 gene binary vector and Agrobacterium-mediated transformation of Chlamydomonas reinhardtii

作　　者：杨泽熵[1] 张雨靖 高文英 吕亚维 王英娟[1] 

机构地区：[1]西部资源与现代生物技术省部共建教育部重点实验室/陕西省生物重点实验室/西北大学生命科学学院,陕西西安710069

出　　处：《西北大学学报（自然科学版）》2017年第1期75-81,共7页Journal of Northwest University（Natural Science Edition）

基　　金：国家自然科学基金资助项目(31270411;31572665);陕西省科技厅社会发展攻关计划基金资助项目(2012K12-01-02);陕西省教育厅专项基金资助项目(12JK0827)

摘　　要：甘油3-磷酸酰基转移酶基因(gpd1)可以调控植物脂质代谢中关键酶甘油-3-磷酸脱氢酶(GPDH)的活性,能促进植物脂质合成。本研究以莱茵衣藻FACHB-479为受体,通过克隆酵母gpd1基因,构建莱茵衣藻的农杆菌双元表达载体pRI-Ble-GPD1,首次以农杆菌介导法将携带在农杆菌LBA4404的gpd1基因转移到FACHB-479基因组。在含有10μg/m L博莱霉素的TAP平板筛选得到抗性藻体细胞。经过分子生物学PCR及RTPCR鉴定,获得转基因藻FACHB-GPD。尼罗红染色法鉴定分析FACHB-GPD细胞中油脂含量增高1.3~1.52倍。该研究表明农杆菌介导外源gpd1基因在莱茵衣藻FACHB-479基因组的过表达能增加脂肪酸含量,从而提高其产油能力。Glycerol-3-phosphate acyltransferase gene（gpd1） regulates the activity of the key enzyme glyceraldehyde-3-phosphate dehydrogenase（GPDH） in plant lipid metabolism to promote plant lipid synthesis. In this study,Agrobacterium binary vector pRI-Ble-GPD1,constructed with the gpd1 gene cloned from Yeast,is firstly transferred into the genome of Chlamydomonas reinhardtii FACHB-479 by the Agrobacterium-mediated method. Transferred alga FACHB-GPD is screened by the TAP solid medium containing 10 μg/m L of bleomycin and identified with the molecular method as PCR and RT-PCR. The analysis with Nile red staining shows the oil content in FACHB-GPD cells is 1. 3 ～ 1. 52 times to the wild type FACHB-479. This study shows that exogenous gpd1 gene transferred by Agrobacterium-mediated method expressed in the genome of Chlamydomonas reinhardtii FACHB-479,can increase the fatty acid content,thereby increasing the capacity of oil production.

关 键 词：莱茵衣藻 农杆菌转化法 GPD1 脂质含量 

分 类 号：Q789[生物学—分子生物学]