Construction of gene library of Arthrobacter BT801 and isolation & expression of hydantoinase gene / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 281-285, 2003.
Article
in Chinese
| WPRIM
| ID: wpr-270068
ABSTRACT
Hydantoinase can be widely used in enzymic production of various amino acids. In order to obtain the hydantoinase genes in Arthrobacter BT801, its chromatosomal DNA is isolated and partialy digested with Sau3A I to collect fragments of about 30kb. Then, this fragment is inserted into the Hpa I and Pst I site of cosmid pKC505. The genomic library was thus constructed by packing in vitro with lambda phage package protein and transfecting E. coli DH5alpha. A positive transformant was selected from the library using thin layer chromatography and other methods. A DNA fragment containing complete hydantoinase genes was sequenced by sub-cloning into pUC18. The gene can express active protein under control of its own promoter and T5 promoter in E. coli. The isolation of the gene established foundition for research and application of the hydantoinase.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Arthrobacter
/
Gene Library
/
Chromatography, Thin Layer
/
Promoter Regions, Genetic
/
Cloning, Molecular
/
Bacteriophage lambda
/
Electrophoresis, Agar Gel
/
Electrophoresis, Polyacrylamide Gel
/
Escherichia coli
/
Amidohydrolases
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2003
Type:
Article
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