Construction Of The BAC Library Of Brassica Napus-Chinese Xinjiang Wild Rape (Sinapis Arvensis) Disomic Alien Addition

Cytoplasmic male sterility(CMS) is widely used in cross breeding,which consists of sterile line,restoring line and/or maintainer line.About 70%of Brassica variaties are Cytoplasmic male sterility(CMS) hybrids.Restoring line is one of the most important parts in the CMS technology.If we can get the restore gene,we could study its structure and use it to develop some more restoring lines,for this purpose we constructed a BAC library.The Sinapis arvensis Xinjiang is crucifer,and a wild breed of Sinapis arvensis L.It is good at resist diseases and insects,low content of erucic acid and glucosinolates.We used the disomic alien addition line which consists two chromosomes of Sinapis arvensis Xinjiang wild rape and the Nsa Cytoplasmic male sterility restore gene,and CopyControl^TM pCC1BACTM(Hindâ…¢Cloning-Ready) Vector followed the manuals of Zhang and Perterson to construct a BAC library.We got plugs with good quality and tested enzyme units at 1.2 U Hind/â…¢,and then we reclaimed the big DNA segments to link with the vector.The library consisting of 69120 clones from the analysis of 100 randomly selected BAC clones,the average insert size is estimated at 110kb,94%are 80-230kb,76%are more than 100kb.As reported that the Brassica napus haploid genome size is 1.2×10⁹bp,the library represents about 6.3×haploid genome equivalents. This provides a 99.82%probability of finding any specific gene.At last we make a primary pool and a secondary pool of this library.The results indicate that this BAC library has high quality and high coverage,and is sufficient for target gene isolation.