玉米端粒酶RNA模板基因候选序列的转基因鉴定体系研究  

Study on Transgenic Identification System of Corn Telomerase RNA Template Gene Candidate Sequence

作　　者：郑洁[1] 杨力媛[1] 马国兴[1] 马登旭[1] 刘小川[1] 

机构地区：[1]浙江理工大学生物工程研究所,杭州310018

出　　处：《浙江理工大学学报（自然科学版）》2013年第5期762-765,774,共5页Journal of Zhejiang Sci-Tech University(Natural Sciences)

摘　　要：研究采用pCAMBIA 1301穿梭质粒和LBA 4404菌株,侵染不同品种玉米成熟胚诱导的愈伤组织,以构建玉米端粒酶RNA候选序列的转基因鉴定体系。构建了两个含有玉米端粒酶RNA的pCAMBIA 1301穿梭质粒,一个携带模板区正常的玉米端粒酶RNA候选序列(野生型),另一个携带模板区定点突变处理的端粒酶RNA候选序列(突变型),然后分别侵染甜玉米成熟胚诱导的愈伤组织,也同样分别侵染糯玉米成熟胚诱导的愈伤组织。结果表明:在愈伤组织培养中,甜玉米的愈伤诱导率在40%左右,而糯玉米的愈伤诱导率高达80%;在抗性筛选培养基上已成功获得抗性愈伤组织,经PCR鉴定愈伤组织基因组中分别含有正常和突变的目的片段,这表明成功构建了玉米端粒酶RNA候选序列的转基因鉴定体系,为进一步鉴定相关候选序列提供有效的技术方法。This study uses pCAMBIA 1301 shuttle plasmid and LBA 4404 bacterial stain for dip dyeing of callus induced by mature embryo of different species of corns so as to establish transgenic identification system of corn telomerase RNA candidate sequence; establishes two pCAMBIA 1301 shuttle plasmids containing corn telomerase RNA, one of which carries corn telomerase RNA candidate sequence with normal template area（wild type） and the other of which carries telomerase RNA candidate sequence with site-specific mutagenesis of template area（mutation type） ; then respectively dip-dyes the callus induced by mature embryo of sweet corn and waxy corn. The result shows that the callus induction rate of sweet corn is about 40% and that of waxy corn is 80% in callus culture; in terms of resistance screening culture medium, resistance callus has been successfully obtained; PCR identification shows that callus genome respectively contains normal and mutational target fragment. This means that transgenic identification system of corn telomerase RNA candidate sequence has been successfully established, thus providing effective technique for further identifying relevant candidate sequences.

关 键 词：甜玉米 糯玉米 端粒酶RNA模板基因候选序列 农杆菌介导法 抗性愈伤组织 

分 类 号：Q785[生物学—分子生物学]