Overproduction, purification and crystallization of TaqI restriction endonuclease |
| |
Authors: | F Barany |
| |
Affiliation: | Department of Microbiology, Hearst Microbiology Research Center, Cornell University Medical College, New York, NY 10021. |
| |
Abstract: | Under phoA promoter control, TaqI endonuclease was overproduced to 5% of Escherichia coli cellular proteins. This was achieved by fusing the endonuclease gene to the first four codons of the alkaline phosphatase signal sequence. For maximal overproduction (30% of cellular proteins), a putative 14-bp hairpin within the endonuclease coding sequence was replaced with degenerate codons. In addition, TaqI methylase was required to protect host DNA. The endonuclease was purified in sufficient amounts for crystallization. |
| |
Keywords: | |
本文献已被 ScienceDirect 等数据库收录! |
|