棘孢木霉TspyrG基因选择标记转化体系的建立  

Establishment of Transformant System for Trichoderma asperellum Using TspyrG as Selected Marker

作　　者：范莉莉 韦润玲 傅科鹤[1] 刘文涛 时晶 FAN Lili;WEI Runling;FU Kehe;LIU Wentao;SHI Jing(Department of Biology,Nanchang Normal University,Nanchang 330032,China)

机构地区：[1]南昌师范学院生命科学学院,南昌330032

出　　处：《西北农业学报》2024年第10期1974-1981,共8页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：江西省教育厅项目(151252,GJJ161233);国家自然科学基金地区项目(31660020)。

摘　　要：旨在从棘孢木霉Ts93中克隆获得尿嘧啶合成关键酶乳清酸核苷-5′-磷酸脱羧酶基因(TspyrG),该基因无内含子,开放式阅读框大小为1140 bp,编码1个379氨基酸的蛋白。通过农杆菌介导技术,获得棘孢木霉Ts93 TspyrG基因缺失突变株ΔTspyrG,突变株在不含尿嘧啶的培养基上无法生长。同时,以PCAMBIA1300质粒为骨架,构建转化载体1300pyrG。以ΔTspyrG突变株为受体菌,成功构建以尿嘧啶营养缺陷为选择标记的木霉菌农杆菌转化系统。对绿色荧光蛋白(eGFP)的转化试验表明,该系统转化效率与抗生素选择标记转化系统无明显差异。In this study,we cloned a TspyrG gene,which encodes orotidine-5′-phosphate decarboxylase,an essential enzyme in pyrimidine biosynthesis in T.asperellum Ts93 strain.The TspyrG gene,which does not contain introns,comprises an open reading frame of 1140 codons and encode a 379 aa protein.The ΔtspyrG mutants were obtained via Agrobacterium-mediated transformation,which were unable to grow without the uracil.And the transformant plasmid 1300pyrG was constructed successfully based on the plasmid PCAMBIA1300.Agrobacterium transformation system with uracil auxotroph maker was successfully constructed using ΔTspyrG as recipient strain.Transformant of eGFP showed no significant difference between auxotrophic selection marker and antibiotic selection marker in the transformant system.

关 键 词：棘孢木霉 营养缺陷型 农杆菌转化系统 

分 类 号：S476[农业科学—农业昆虫与害虫防治]