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Updated: Jul 2, 2026

Defining Substrate Specificities for Lipase and Phospholipase Candidates
Published on: November 23, 2016
Cloning, sequence analysis and expression of bacterial lipase-coding DNA fragments from environment in Escherichia
Zhaoxin Fan1, Changwu Yue, Yang Tang
1College of Life Science, Sichuan University, Sichuan Key Laboratory of Molecular Biology & Biotechnology, Chengdu, 610064, China. fanzhaoxin@sohu.com
Abstract:
Thirteen pairs of primers were designed, synthesized and used to clone the whole coding sequences or mature peptide-coding sequences of lipases. Bacteria producing extracellular lipases were enriched for the extraction of total DNAs. Eight fragments with 500-1,200 bp in length were obtained by using touchdown PCR and sequenced. Five of them were found to be lipase-coding DNAs. One fragment called BL9 that was 95.9% similar to a coding sequence of putative lipase. This lipase contained a Gly-His-Ser-Met-Gly motif which is matched to the consensus Gly-X-Ser-X-Gly conserved among lipolytic enzymes. The BL9 DNA fragment was inserted into the expression vector pET32a(+) of Escherichia coli. A functional product was yielded in the supernatant and produced a hydrolyzed zone on the tributyrin agar.
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