BADH/pepB双价基因无选择标记表达载体转化苜蓿的初步研究  

Research on Transforming BADH and pepB Bivalent Gene with Marker-free Expression Vector into Alfalfa

作　　者：杨金慧[1] 王丕武[1] 曲静[2] 王巍[2] 王俊玲[2] 

机构地区：[1]吉林农业大学农学院,吉林长春130118 [2]吉林农业大学生命科学院,吉林长春130118

出　　处：《中国草地学报》2009年第4期20-23,共4页Chinese Journal of Grassland

基　　金：吉林省科技发展计划项目(20040520)

摘　　要：选用公农1号紫花苜蓿，以5～7d苗龄的无菌子叶为外植体，通过农杆菌介导法将含有甜菜碱醛脱氢酶（BADH）和酸性蛋白酶（pepB）双价基因的无选择标记表达载体导入苜蓿子叶中，并用含有Na2CO3和NaHCO3碱性盐溶液的培养基进行筛选，得到抗盐碱转化植株。碱性盐浓度48mmol／L宜于子叶愈伤诱导，有利于转化植株的获得。对转化植株进行PCR检测，初步证明目的基因序列已经整合到苜蓿基因组中。移栽成活的13棵碱性盐抗性植株经PCR检测有3棵植株呈阳性。The cotyledons of alfalfa were selected as explants for genetic transformation and were cultvated with Agrobacterium tumefaciens strain EHA105 harboring marker-free expression vector containing BADH and pepB binary genes. Regenerated plants were obtained in the selection media supplemented with 48mmol/L Na2CO3 and NaHCO3. The genomic DNA of transgenic plants were extracted and analyzed by PCR. Result showed that BADH and pepB genes were transformed into the genome of transgenic alfalfa. 3 plants among 13 plants grown up through selection on medium containing 48mmol/L Na2CO3 and NaHCO3 were assessed by PCR amplification of 1340 bp of pepB gene.

关 键 词：无选择标记表达载体 BADH pepB 转化 苜蓿 

分 类 号：Q78[生物学—分子生物学]