Cloning of Laccase Gene Fragments and Detection of Laccase Activity in Setosphaeria turcica

doi:10.7668/hbnxb.2011.02.017

Abstract

Abstract: The object of this paper is to examine the existence of laccase in Setosphaeria turcica and influencing factors on enzyme activity.The laccase activity of S.turcica was tested by spectrophotometer under 420 nm with ABTS(2,2：azino-bis(3-ethyl-2,3·dihydro-1 3 benzthiazole-6-sulfonate))as the substrate,and the influence of Cu“concentration and reaction pH value on the enzyme activity were studied.A pair of degenerate primers was de- signed according to the conserved Cu2+domain of other fungal laccases and homologous fragment of laccase gene was cloned by polymerase chain reaction with cDNA as template.This study demonstrated that there was laccase in S. turcica and the laccase activity was related to copper concentration and pH.The results showed that optimum condi- tions were pH2.6 and 250 trmol/L CuS04 respectively.In our research,a 929 bp fragment was obtained from S.turcica.,named as StLacl,which shared over 60%identity with other fungus laccase and contained Cu-oxidise superfam- ily.This work is helpful to study and understand laccase function of S.turcica.

Key words: Setosphaeria turcica, Laccase, ABTS, Clone

CLC Number:

S432.1

CAO Zhi-yan, ZANG Jin-ping, JIA Hui, LI Xu-guang, WANG Ping. Cloning of Laccase Gene Fragments and Detection of Laccase Activity in Setosphaeria turcica[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2011, 26(2): 76-80. doi: 10.7668/hbnxb.2011.02.017.

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