结球甘蓝cDNA文库的构建和鉴定

cDNA Library Construction and Identification of Cabbage

构建了结球甘蓝cDNA文库。文库的宿主菌为E .coliXL1 Blue。对文库的部分特性进行研究 ,滴度为 2× 10 8～ 4× 10 8pfu/mL ,插入片段的大小均在 5 0 0bp以上 ,以 1kb以上的片段占大多数 ,重组体的重组率为 95 % ,用抗病基因片段作探针进行噬菌体原位杂交 ,杂交信号明显。对文库进行了扩增、保存。

A cDNA library of cabbage was constructed and an expression plasmid introduced into E.coli XL1 Blue was used as cloning vector. The titer of cDNA library is 2×10 8-4×10 8 pfu/mL; most of the fragments are more than 500bp, the recombinant rate of the library is about 95%. Probed with resistant gene fragment ,phage in situ hybridization ,the hybridization signal is clear, which shows this library is good ,and the library had been amplified and stored.