pyrG Gene Cloning and Establishment of Homologous Transformation System for Aspergillus oryzae

doi:10.7506/spkx1002-6630-201011043

FOOD SCIENCE ›› 2010, Vol. 31 ›› Issue (11): 202-205.doi: 10.7506/spkx1002-6630-201011043

• Bioengineering • Previous Articles Next Articles

pyrG Gene Cloning and Establishment of Homologous Transformation System for Aspergillus oryzae

WANG Jin-liang，CHEN Hong-wen*

Key Laboratory of Industrial Biotechnology of Fujian Province University, Huaqiao University, Quanzhou 362021, China

Received:2009-10-14 Online:2010-06-01 Published:2010-12-29
Contact: CHEN Hong-wen E-mail:chenhw@hqu.edu.cn

Abstract

Abstract:

The present paper reports the establishment of homologous transformation system for Aspergillus oryzae based on pyrG gene as the selective marker. pyrG gene was amplified by PCR with A. oryzae genome as a template and connected with pMD-18T vector. The product was transformed into E. coli DH 5α. Recombinant plasmid pMD-pyrG was verified according to blue-white screening, PCR fast screening, enzyme digestion and sequence identification. Sequence analysis exhibited 99.9 % homology with the pyrG gene of A. oryzae KNB 616. The deduced homology for amino acids was 99.6 %. Transformation of A. oryzae pyrG－ mutant with recombinant plasmid pMD-pyrG by PEG/CaCl2 method could result in its genetic transformation to become pyrG+ mutant. The establishment of homologous transformation system for A. oryzae makes it possible to introduce exogenous gene and analyze its function in the future.

Key words: recombinant plasmid, cloning, homologous transformation system

CLC Number:

Q785

WANG Jin-liang，CHEN Hong-wen*. pyrG Gene Cloning and Establishment of Homologous Transformation System for Aspergillus oryzae[J]. FOOD SCIENCE, 2010, 31(11): 202-205.

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pyrG Gene Cloning and Establishment of Homologous Transformation System for Aspergillus oryzae

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