Li Jin, Yang Gong She, Chen Chuang Fu, Liu Hai Bo, Ma Run Lin, Wang Jin Fu
College of Animal Science & Technology, Shihezi University, Shihezi.
Fen Zi Xi Bao Sheng Wu Xue Bao. 2006 Dec;39(6):489-94.
In this study, we used our special regional fostering breed Xinjiang fine-fleece sheep's genome to construct its genome library. The average size of insert fragments of the library was 133 kb. At the same time 92.5% clones' insert fragments of the library were larger than 100 kb and some larger than 300 kb. If we supposed sheep's genome contenting 3 x 10(6) kilo-base, on the basis of the average insert fragment was 133 kb the library covered 8 times genome of Xinjiang fine-fleece sheep. The probability of the tagged fragment being screened from the library was 98.208%. To prove the BAC library had been the better rate of coverage,four molecular markers: DMB_EX2, MCMA36, CP73 and BM1258 located to MHC gene of chromosome 20 near region in xinjiang fine-fleece sheep had been screened positive clones from the constructed 2 times genome library PCR screening system and the average positive clones was 1.5. The screening result showed that the constructed genome library was fairly closed to the 8 times genome coverage and had no erroneous tendency, which made the library being of the utmost useful resource for studying functional gene, position cloning and improving the genome physical map of sheep.
在本研究中,我们利用我国特有的地方培育品种新疆细毛羊的基因组构建其基因组文库。该文库插入片段的平均大小为133 kb。同时,文库中92.5%的克隆插入片段大于100 kb,部分大于300 kb。若假定绵羊基因组大小为3×10(6)千碱基,基于平均插入片段为133 kb,该文库覆盖了新疆细毛羊基因组的8倍。从文库中筛选到标记片段的概率为98.208%。为证明BAC文库具有较好的覆盖率,从构建的2倍基因组文库PCR筛选体系中,对位于新疆细毛羊20号染色体MHC基因附近区域的4个分子标记:DMB_EX2、MCMA36、CP73和BM1258进行阳性克隆筛选,平均阳性克隆数为1.5个。筛选结果表明,构建的基因组文库接近8倍基因组覆盖率,且无错误倾向,这使得该文库成为研究功能基因、定位克隆及完善绵羊基因组物理图谱的极为有用的资源。