THE ARYLDIALKYLPHOSPHATASE-ENCODING GENE-ADPB FROM NOCARDIA SP STRAIN-B-1 - CLONING, SEQUENCING AND EXPRESSION IN ESCHERICHIA-COLI

被引:28
作者
MULBRY, WW
机构
[1] Pesticide Degradation Laboratory, ARS, USDA, Beltsville
关键词
PARATHION HYDROLASE; BIODEGRADATION; PARAOXONASE; ORGANOPHOSPHATE; PESTICIDE; HYDROLASE;
D O I
10.1016/0378-1119(92)90174-N
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
Using degenerate oligodeoxyribonucleotides (oligos) derived from the N-terminal sequence of an aryldialkylphosphatase (ADPase) from Nocardia sp. strain B-1, an amplification reaction was used to isolate a DNA segment containing a 57-bp fragment from the adpB gene. Based on the nucleotide (nt) sequence of this fragment, a nondegenerate oligo was synthesized and used to screen a subgenomic library of strain B-1 DNA for fragments containing adpB. A 3.55-kb PstI fragment containing adpB was cloned into Escherichia coli, and the nt sequence of a 1600-bp region containing adpB was determined. Under control of the lac promoter of pUC19, adpB expression in E. coli cultures was approx. 15-fold higher than in strain B-1 under the native adpB promoter. Comparison of adpB with the Flavobacterium ADPase-encoding gene, opd, revealed no significant homology at the nt or aa levels.
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页码:149 / 153
页数:5
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