Obtainment of Transgenic Tobacco Harboring phbA , phbB and phbC Genes by Twice Transformation  

二次转化获得整合phbA、phbB、phbC基因的转基因烟草(英文)

作　　者：张景昱[1] 叶梁[1] 李丽 宋艳茹[1] 

机构地区：[1]中国科学院植物研究所,北京100093

出　　处：《Acta Botanica Sinica》2001年第1期59-62,共4页Acta Botanica Sinica（植物学报：英文版）

基　　金：SupportedbytheState"863"HighTechnologyProgram ( 10 1_0 4_0 3_0 4) ;theNationalNaturalScienceFoundationofChina ( 5 9673 0 0 8)

摘　　要：To avoid the long time required for conventional sexual crossing, transgenic tobacco (Nicotiana tabacum L.) plants harboring phbA gene (encoding 3_ketothiolase) were used as the target plant for the second transformation mediated by Agrobacterium tumefaciens (Smith et Townsend) Conn LBA4404 containing pZCB which was constructed by linking phbB (encoding acetoacetyl_CoA reductase), phbC (encoding PHB synthase) and ctp sequence to pBIB_HYG under the control of CaMV 35S promoter. The hygromycin resistant transformants were morphologically normal and stable integration of phbB and phbC was confirmed by PCR and PCR_Southern. Moreover, RT_PCR_DNA hybridization analysis showed that 6.67% of the transformed tobacco plants could express both phbB and phbC at transcriptional level.将携有导肽序列的phbB(编码乙酰乙酰CoA还原酶 )和phbC(编码PHB合酶 )连入pBIB_HYG得到组成型表达载体pZCB ,用冻融法转入根癌土壤杆菌 (Agrobacteriumtumefaciens (SmithetTownsend)Conn)并由其介导转化已整合且表达phbA(编码 3_酮硫裂解酶 )基因并具有卡那霉素抗性的转基因烟草 (NicotianatabacumL .)。通过二次转化可避开传统杂交育种 ,在 5个月内获得整合PHB合成所需 3个基因的转基因烟草。所获转基因植株表型正常 ,经PCR、PCR_Southern、RT_PCR_DNA杂交检测确定有 5 0株烟草稳定整合phbB、phbC基因 ,其中 6 .6

关 键 词：poly (3_hydroxybutyrate) (PHB) twice transformation transgenic tobacco 

分 类 号：Q943.2[生物学—植物学]