黑曲霉转化中作为同源标记的sC的克隆与应用

Cloning and use of sC as homologous marker for Aspergillus niger transformation.

作者信息

Varadarajalu Lakshmi P, Punekar Narayan S

机构信息

Biotechnology Group, School of Biosciences and Bioengineering, Indian Institute of Technology, Bombay, India.

出版信息

J Microbiol Methods. 2005 May;61(2):219-24. doi: 10.1016/j.mimet.2004.11.022. Epub 2004 Dec 19.

DOI:10.1016/j.mimet.2004.11.022

PMID:15722148

Abstract

The sC sequence from Aspergillus niger was cloned and developed into a homologous marker system for genetic transformation. The coding region of the sC gene amplified by PCR from the A. niger genome was provided with Aspergillus nidulans expression signals (gpdA promoter and trpC terminator). This chimeric construct was used to successfully transform a spontaneous sC- isolate of A. niger to prototrophy. The transformants analyzed by Southern analysis showed integration of multiple copies of the transforming DNA. They also exhibited much higher ATP sulfurylase activity than the wild-type A. niger strain reinforcing the molecular data. This demonstrates the usefulness of the sCniger construct, driven by PgpdA, as a marker for A. niger transformation.

摘要

克隆了来自黑曲霉的sC序列，并将其开发成用于遗传转化的同源标记系统。通过PCR从黑曲霉基因组中扩增出的sC基因编码区带有构巢曲霉表达信号（gpdA启动子和trpC终止子）。该嵌合构建体用于成功地将黑曲霉的自发sC-分离株转化为原养型。通过Southern分析的转化体显示出转化DNA的多拷贝整合。它们还表现出比野生型黑曲霉菌株更高的ATP硫酸化酶活性，这加强了分子数据。这证明了由PgpdA驱动的sC黑曲霉构建体作为黑曲霉转化标记的有用性。