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Ti质粒介导的磷酸烯醇式丙酮酸羧化酶cDNA转化烟草植株

董龙英, 凌俊, 王朝明, 施教耐

中科院上海植物生理研究所. 上海 200032

摘要

将玉米C_(4)-磷酸烯醇式丙酮酸羧化酶cDNA亚克隆至穿梭质粒pBin19, 通过农杆菌Ti质粒(LBA4404)介导的叶圆片共培养法将其转入C_(3)植物烟草中。在获得的抗 性转化植株中, 80％具有较强的NPTⅡ报道基因表达。Southern杂交表明C_(4)-PEP羧化酶cDNA已被整合到了烟草核基因组中。

关键词： 农杆菌; Ti质粒; PEP羧化酶cDNA; 烟草; 基因转移

Ti plasmid-mediated transformation of tobacco with C_(4)-PEPCase cDNA from Zea mays

Dong Longying, Ling Jun, Wang Chaoming, Shi Jiaonai

Shanghai Institute of Plant Physiology, Chinese Academy of Sciences. Shanghai 200032

Abstract

A2.0kb fragement of C_(4)-PEP-Case cDNA derived from a maize PEP-Case cDNA clone pPEP3055 was subcloned into binary vector pBin 19, and transconjugated into Agrobacterium tume-faciens) retaining vir helper plasmid LBA 4404 by means of ‘triparental mating’. The the engineering plasmid pBPC2 harbouring C_(4)-PEPCase cDNA was transformed into the leaf disc of tobacco by co-culture with transconiugated A. tumefaciens). Antibiotic screening yielded a batch of transformants resistant to kanamycin and the regenerated transgenic tobacco was obtained.

Key words: Agrobacterium tumefaciens;Ti plasmid;PEPCase cDNA;Tobacco;Gene transformation

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引用本文：

董龙英, 凌俊, 王朝明, 施教耐. Ti质粒介导的磷酸烯醇式丙酮酸羧化酶cDNA转化烟草植株[J]. 生理学报 1995; 47 (3): .

Dong Longying, Ling Jun, Wang Chaoming, Shi Jiaonai. Ti plasmid-mediated transformation of tobacco with C_(4)-PEPCase cDNA from Zea mays. Acta Physiol Sin 1995; 47 (3): (in Chinese with English abstract).