China Animal Husbandry & Veterinary Medicine ›› 2007, Vol. 34 ›› Issue (10): 53-55.

• 生物技术 • Previous Articles     Next Articles

Construction of Fosmid Library in Domestic Pig

LI Bing1,YAN Shouqing1,SUN Jinhai1,2
   

  1. 1.College of Animal Science and Veterinary Medicine，Jilin University，Changchun 130062，China;2.College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2007-10-20 Published:2007-10-20

Abstract: Genomic DNA prepared for library construction was digested by proteina
se K and extracted by phenol. The DNA was randomly sheared and endrepaired by T4 DNA Polymerase and T4 Polynucleotide Kinase.25~40 kb fragment was recovered by 1% agarose gel electrophoresis and ligated with pCC1FOS Vector. Recombinant molecules were packaged in vitro with the Phage Packaging Extract, then transformedinto E coli EPI300. A fosmid genomic library of domestic pig was successfully constructed with the titer of 8.50×109 CFU/ml in which the insert fragment size was about 25 kb.Positive clones were achieved by in stitu hybridization with MC1satellite DNA as probe.It was indicated that the library was of high quality which can be used to screen new satellite DNA in domestic pig.

Key words: domestic pig; fosmid library; satellite DNA

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