›› 2010, Vol. 37 ›› Issue (3): 112-114.

• 生物技术 • Previous Articles     Next Articles

Construction and Analysis of a Full-length cDNA Library of Ascosphaera apis

ZHAO Hong-xia1，2,LI Jiang-hong1,LIANG Qin1,LUO Yue-xiong2,ZHANG Xue-feng2   

  1. (1.College of Bee Science, Fujian Agriculture and Forestry University, Fuzhou 350002,China;2.Guangdong Entomological Institute, Guangzhou 510260,China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-03-20 Published:2010-03-20
  • Contact: LIANG Qin

Abstract: In this study，the total RNA of Ascosphaera apis was extracted by using TRIzol reagent. The Oligo reverse transcriptase was used to synthesize and anchor the first strand Cdna.Following the production of the first reverse transciption,we synthesize the second Cdna. The production of cDNA was separated by classes and packed outside, then we could get the cDNA original library of Ascosphaera apis.The unamplified cDNA library contaied 1.6×106 PFU/mL independent clones. The titer of the amplified library was 1.5×109 PFU/mL. Taking suitable amplified library to dilute and litter the flat, picking single plaques and then using common primer M13± to amplify，the recombination clones with an average insert size of 0.5 to 2 kb was higher than 90%. The construction of this library will provide a platform for the research of invasion mechanism and prevention.

Key words: Ascosphaera apis; full-length cDNA library; lambda phage

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