作者:
摘要:<正> Bacillus thuringiensis (Bt) strain C002 contains crylAa, cry2Ab, crylCa insecticidal crystal genes and an unkown gene cryX, among which crylCa is located in a 6 -9 kb EcoR I fragment of the chromosomal DNA. The total DNA and the plasmids DNA libraries of C002 were constructed in Bt-E. coli shuttle plasmid pHT315 by inserting 6 - 9 kb chromosomal and plasmid DNA fragments prepared respectively with EcoR I complete and 5au3A I partial digestion. On the basis of every 50 transformants pooled together from 5-10 tubes, the pools containing about 2 000 transformants from the plasmids DNA library and 400 transformants from the total DNA library were rapidly screened by PCR-RFLP. Clones containing crylAa, cryX, crylCa , and cry2Ab were isolated and named as pHT-1Aa, pHT-X, pHT-1Ca and pHT-2Ab respectively. Restriction analysis indicated that pHT-1Aa, pHT-1Ca and pHT-2Ab had the typical physical map of the homologous cry genes. Furthermore, each plasmid was transferred into Bt acrystalliferous strain cryB- by el
关键词:Bacillus thuringiensis;DNA library;PCR-RFLP;cry genes;Gene cloning;
文献注录:. Cloning of Bt cry Genes by Rapid Screening of DNA Libraries with PCR-RFLP [J]. Agricultural Sciences in China. 2003, 02: 18-22.
报/刊名:《Agricultural Sciences in China》,发表于2003 / 第 02 期。
文献类型: [J] (文献级别:核心刊物)
页码:第 18-22 页 / 共5 页