Construction Of BAC Library Of Xinjiang Wild Rape And Study On Screening Of A Single Chromosome Clone

Rape is one of the important oil crops. Xinjiang wild rape is a wild mustard (Sinapis arvensis) found in a natural population of Xinjiang, China, it has strong growth potential, strong resistance to pests and diseases, and strong resistance to Sclerotinia sclerotiorum, as well as sterile cytoplasm, so it is a fine breeding of wild rapeseed germplasm. Brassica-Xinjiang wild rape disomic addition lines (B. napus-S. arvensis disomic alien addition line) is a new rapeseed restorer line which is obtained through protoplast fusion between Xinjiang wild rape 18 and ZhongShuang 4 (Brassica napus).and continuously backcross four times.The disomic addition lines have a pair of chromosomes from Xinjiang wild rape, which carrying the fertility restorer gene. Cloning of the restorer gene is to explore recovery mechanism, and cultivate good transgenic rapeseed restorer lines, It has important scientific significance and production value. The study has constructed wild rape genomic BAC library and studied BAC clones of Xinjiang wild rape chromosome from Brassica napus-Xinjiang wild rape disomic addition lines of BAC library. The results are as follows:Construction of Xinjiang wild rape genomic BAC library:With low melting point agarose embedded nuclei,high molecular weight DNA were prepared, then digested by HindⅢ, through electro elution,100-300kb large fragment DNA were recovered and linked with pCC1BACHTM vector, the connection was desalted and concentrated with 0.025μm dialysis membrane suspended in 10% PEG8000 on ice, then transfered into E.coil EP1300 competent cells. Xinjiang wild rape genomic BAC library had been constructed, and stored in 38 384 well plates. Randomly selected 100 clones, digested with NotI, tested insert size with PFGE, the library has 14,592 clones, with the insert size of 50-250kb, average insert of 105kb, covering the genome of Xinjiang wild rape 4.3 times.Screening specific BAC clones from wild rape chromosome:Xinjiang wild rape genomic DNA as a probe, directly hybridized colony in situ to the Brassica napus-Xinjiang wild rape disomic addition lines of clone BAC library to obtain a specific chromosome of wild rape sources of BAC clones, but get no desired results, the right dose of Brassica napus genome DNA for blocking will be carried on for the further correlation research.