ADVANCES IN POLAR SCIENCE ›› 2013, Vol. 25 ›› Issue (3): 249-256.DOI: 10.3724/SP.J.2013.00249

Previous Articles     Next Articles

CLONING OF THE β-GALACTOSIDASE GENES FROM MARINOMONAS SP. BSI20584 by hiTAIL-PCR

Zhou Lili1,2,Chen Ruiqin1,2,Chen Xiulan3,Zeng Yinxin2,Chen Bo1,2   

  1. 1School of Biotechnology of East China University of Science and Technology, Shanghai 200237,China; 2Polar Research Institute of China,Key Laboratory for Polar Science, SOA, Shanghai 200136,China;3State Key Laboratory of Microbial Technology, Jinan 250100
  • Received:2012-12-19 Revised:2013-01-15 Online:2013-09-30 Published:2013-09-30

Abstract: A complete gene of a β-galactosidase was isolated from Marinomonas sp. BSi20584 by hiTAIL-PCR(high-efficiency thermal asymmetric interlaced). A pair of primes were designed according to online conserved regions in other β-galactosidases.The conversed fragment of β-galactosidase was gained by PCR. The N-terminal DNA sequence was amplified by PCR using primers based on N-terminal amino acid sequence of the purified enzyme. Upstream and downstream of sequences already obtained and named nbs were gained  by hiTAIL-PCR. In the end , the whole gene was amplified by primers designed according to the assembly DNA sequences. In this article the β-galactosidase gene was successfully isolated and was 1971bp. NCBI blast indicated it was a new member of glycosylhydrolase family 42.

Key words: β-galactosidase, hiTAIL-PCR, gene walking