Ahmed S S, Gong Z-H, Ji J-J, Yin Y-X, Xiao H-J, Khan M A, Rehman A, Ahmad I
College of Horticulture, Northwest A&F University, Shaanxi, Yangling, P.R. China.
Genet Mol Res. 2012 Aug 31;11(3):3091-104. doi: 10.4238/2012.August.31.7.
Molecular chaperones of plasmid pBI121 carrying CaMV35S promoter and a nucleotide sequence of plasmid pBI221 were inserted into plasmid pCAMBIA2300 to construct an intermediate vector: pVBG2307. This novel vector pVBG2307 contains a greatly expanded multiple cloning site with an adjacent imported CaMV35S promoter sequence. This vector allows controlled transformation of DNA in both Escherichia coli and Agrobacterium tumefaciens. Cloned PG, orf456, ipt genes and E8, a fruiting promoter, were amplified by PCR of cDNA libraries of Capsicum annum and Lycopersicon esculentum and were then transferred into vector pVBG2307. The viability of this vector was demonstrated, as it regulated PG, orf456, ipt and E8 genes in E. coli and could be transferred into Agrobacterium strain EHA105-4.
将携带花椰菜花叶病毒35S启动子的质粒pBI121的分子伴侣和质粒pBI221的核苷酸序列插入质粒pCAMBIA2300中，构建中间载体：pVBG2307。这种新型载体pVBG2307包含一个大大扩展的多克隆位点以及相邻的导入花椰菜花叶病毒35S启动子序列。该载体允许在大肠杆菌和根癌农杆菌中对DNA进行可控转化。通过对辣椒和番茄的cDNA文库进行PCR扩增克隆的PG、orf456、ipt基因以及果实成熟启动子E8，然后将它们转移到载体pVBG2307中。该载体的活性得到了证明，因为它在大肠杆菌中调控PG、orf456、ipt和E8基因，并且可以转移到根癌农杆菌菌株EHA105 - 4中。
