同济大学学报（医学版）2001，Vol.22Issue(3)：1-3,3.
黑曲霉菌为宿主菌的重组表达性质粒的构建
Construction of Recombinant Expression Plasmid for Aspergillus niger
刘钟滨 1DavidJJeenes2
作者信息
- 1. 同济大学医学院微生物与免疫学教研室
- 2. Institute of Food Research, Norwich NR4 7UA, UK
- 折叠
摘要
Abstract
Objective To establish a protein expression system using pyrG gene as a selection marker and Aspergillus niger (A.niger) as a host.Methods A 9.8kb DNA fragment was isolated after screen of λphage genome library of A.niger ATCC 12049 with part sequence of pyrG gene as a probe which was obtained by PCR. Then a 2.3 kb fragment containing pyrG gene was subcloned into an expression plasmid PIGF. The whole pyrG gene was sequenced and analysed. Transformation was carried out to transfer the recombinant plasmid into A.niger M54, a pyrG mutant of A, niger.Results The pyrG gene was isolated from genome library of A.niger ATCC 12049 and the recombinant plasmid containing pyrG gene was constructed, getting a recombinant plasmid pYG1.2. Sequence analysis showed the pyrG gene cloned was 93.9% homology with that of A. niger L112 and 98.9% homology with the deduced amino acid sequence of it. Transformation of A. niger M54 resulted in stable Pyr+ transformants. Conclusion The results indicated that the recombinant expression plasmid contaning pyrG gene as a selection marker and A.niger as a gene engineering host had been successfully established.关键词
重组质粒/构建/黑曲霉菌引用本文复制引用
刘钟滨,DavidJJeenes..黑曲霉菌为宿主菌的重组表达性质粒的构建[J].同济大学学报（医学版）,2001,22(3):1-3,3.
