ipt与PAPⅡ基因在烟草中共表达减轻PAPⅡ细胞毒性的初步研究  

A Primary Study on Alleviation of PAPⅡCytotoxicity by co-Expression of ipt Gene with PAPⅡin Tobacco

作　　者：殷娴[1,2] 李朝霞[1,3] 吴国智 程伟[1] 王晴 陈正华 张海燕 肖尊安[1] 

机构地区：[1]北京师范大学生命科学学院,北京100875 [2]甘肃亚盛集团博士后科研工作站北京分站,北京100101 [3]中国科学院遗传与发育研究所,北京100101 [4]天津市园林花圃,天津300112 [5]甘肃亚盛实业(集团)股份有限公司,甘肃兰州730030

出　　处：《激光生物学报》2011年第2期254-260,共7页Acta Laser Biology Sinica

基　　金：转基因生物新品种培育重大专项(2009ZX08009-089B)

摘　　要：利用农杆菌介导方法,分别将PAPⅡ单基因、PAPⅡ基因和异戊烯基转移酶基因(ipt)共表达的双基因(PAPⅡ-ipt)导入烟草品种K326叶细胞中,半定量RT-PCR分析了转基因植株中PAPⅡ、ipt和核糖体蛋白大亚基基因(RPL3A)的表达,并进行了TMV攻毒实验。结果表明:双基因PAPⅡ-ipt对烟草的转化频率高达45.55%,为单基因PAPⅡ的转化频率(13.3%)的3.4倍;50%的转基因植株没有检测到转基因的表达,PAPⅡ-ipt表达的转基因植株中RPL3A基因的表达水平为PAPⅡ表达植株中的1.25倍,且生长发育正常,对TMV病毒的抗性有较大程度的提高。因此,ipt与PAPs基因共表达可作为克服PAPs细胞毒性的有效途径。The gene PAP Ⅱ and the co-expressed genes of PAP Ⅱ -ipt （ isopentenyl transferase） were introduced into mesophyll cells of Nicotiana tabacum L. var. K326, respectively, mediated by Agrobacterium tumefaciens. The expression levels of the genes such as PAP Ⅱ , ipt and RPL3A （ Ribosomal Protein L3 ） in transgenic plants were determined by semi-quantitative RT-PCR, and the TMV challenge experiment was conducted for the transgenic plants. The results showed that transformation frequency of the double genes PAP Ⅱ -ipt reached 45.55 % , 3.4 times as much as that of PAP Ⅱ gene alone, transcription of the trans-genes was not determined in 50 % of the transgenic plants, and the expression level of RPL3A in transgenic plants with expression of the double genes was 1.25 times as much as that in transgenic plants with expression of the single PAP Ⅱ gene. Moreover, the transgenic plants with co-expression of the PAP Ⅱ and ipt genes had normal phenotype and fertility, as well as exhibited higher degree of resistance to the TMV. Thus, it might be suggested that the co-expression of ipt and PAP Ⅱ genes is an effective solution to reduce PAPs cytotocixity.

关 键 词：美洲商陆抗病毒蛋白 异戊烯基转移酶 细胞毒性 核糖体蛋白L3 

分 类 号：Q78[生物学—分子生物学]