Construction of metagenomic DNA library of deep-sea sediments and screening of amylase

HUANG Yali; ZHANG Jiong; CAO Lixiang; TAN Hongming; LU Yongjun; ZHOU Shining

doi:10.3969/j.issn.0253-4193.2013.01.016

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HUANG Yali, ZHANG Jiong, CAO Lixiang, TAN Hongming, LU Yongjun, ZHOU Shining. Construction of metagenomic DNA library of deep-sea sediments and screening of amylase[J]. Haiyang Xuebao, 2013, 35(1): 144-148. doi: 10.3969/j.issn.0253-4193.2013.01.016

Citation:

HUANG Yali, ZHANG Jiong, CAO Lixiang, TAN Hongming, LU Yongjun, ZHOU Shining. Construction of metagenomic DNA library of deep-sea sediments and screening of amylase[J]. Haiyang Xuebao, 2013, 35(1): 144-148. doi: 10.3969/j.issn.0253-4193.2013.01.016

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Construction of metagenomic DNA library of deep-sea sediments and screening of amylase

doi: 10.3969/j.issn.0253-4193.2013.01.016

1.
School of Fundamental Medical Science, Guangzhou University of Chinese Medicine, Guangzhou 510006, China;State Key Laboratory for Biocontrol, Department of Biochemistry, College of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China
2.
Beijing Genomics Institute at Shenzheng, Shenzhen 518083, China
3.
State Key Laboratory for Biocontrol, Department of Biochemistry, College of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China

Received Date: 2011-06-15
Rev Recd Date: 2012-08-13

Abstract

Abstract

Deep sea environment is usually characterized by extremely high salinity, high pressure, high/low temperature and darkness. The physicochemical features ensured that may exist in microorganisms unique metabolic and physiological activities, molecular and cellular structure to adapt to these extreme conditions. Therefore, the exploration of biological and function diversity of microorganisms in deep sea are expected to yield novel metabolites. To access to the microbial genetic resources of deep-sea sediment with a culture-independent approach, The sediment DNA was extracted from deep-sea sediments of the South China Sea and cloned into fosmid vector generating a library of 39 600 clones with inserts of 24-45 kb and an average insert size of 33 kb. The metagenomic library represented about 1 320 Mbp of genomic DNA in deep-sea sediments. The library was screened for amylase activity and three recombinant clones showed amylolytic activity. The clone, amy7, with lower optimal temperature was further analyzed. Subsequent library of insert DNA of amy7 was constructed and a positive subclone, designated as amy7-4, was screened. Sequencing of the clone revealed that it contains a 3 291 bp fragment. An open reading frame of 1 920 bp was identified from the fragment and showed high identity with genes of glycosidase from various organisms.
- deep-sea sediment,
- fosmid metagenomic library,
- marine microorganisms,
- amylase

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References(13)

References

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