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Journal of Animal and Veterinary Advances
Year: 2014  |  Volume: 13  |  Issue: 11  |  Page No.: 682 - 687

Construction and Analysis of Rumen Bacterial Artificial Chromosome Library from a Cashmere Goat Rumen Microflora

Mei-Li Zhang and Yu-Lin Chen    

Abstract: A Bacterial Artificial Chromosome Library of rumen microorganism from Shanbei Cashmere goat was constructed to study the potential of the ruminant’s ecosystem and the library storage capacity, stability and the functionality was also analyzed. The high molecular weight DNA was extracted and purified directly from rumen samples in the study by using embedding technology and pulsed field gel electrophoresis approaches. After digestion with HindIII, DNA fragments ranging from 50-150 kb was collected and ligated to pCC1BAC vector. The ligation mixture was transformed into E. coli TransforMax EPI300. The rumen microorganism BAC library contains about 14,400 clones and the average insert size of BAC clones was estimated to be 74.1 kb, mostly ranging from 50-90 kb. Based on the data mentioned above, the capacity of the BAC library was about 1,067 Mb, the frequency of clones without inserts was <2%. The BAC clones show good stability after cultured for 100 generations. RFLP analysis showed a great diversity of the insert fragments. Several BAC clones with activity of amylase had been screened from the BAC library. Eight clones with amylase activity were acquired. It was suggested that the constructed BAC library had good storage capacity, stability and functionality and could be used for further research on exploration of enzyme resource and rumen microflora of Cashmere goat.

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