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TaqI限制性内切酶的过量生产、纯化及结晶

Overproduction, purification and crystallization of TaqI restriction endonuclease.

作者信息

Barany F

机构信息

Department of Microbiology, Hearst Microbiology Research Center, Cornell University Medical College, New York, NY 10021.

出版信息

Gene. 1988 May 30;65(2):167-77. doi: 10.1016/0378-1119(88)90453-2.

DOI:10.1016/0378-1119(88)90453-2
PMID:2842231
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.

摘要

在phoA启动子的控制下，TaqI核酸内切酶的产量过量至大肠杆菌细胞蛋白质的5%。这是通过将核酸内切酶基因与碱性磷酸酶信号序列的前四个密码子融合来实现的。为了实现最大过量表达（细胞蛋白质的30%），核酸内切酶编码序列中的一个推定的14碱基对发夹结构被简并密码子取代。此外，需要TaqI甲基化酶来保护宿主DNA。纯化得到了足够用于结晶的核酸内切酶。

相似文献

1
Overproduction, purification and crystallization of TaqI restriction endonuclease.TaqI限制性内切酶的过量生产、纯化及结晶
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引用本文的文献

1
Genetic organization and molecular analysis of the EcoVIII restriction-modification system of Escherichia coli E1585-68 and its comparison with isospecific homologs.大肠杆菌E1585-68的EcoVIII限制修饰系统的遗传组织与分子分析及其与同特异性同源物的比较。
Appl Environ Microbiol. 2003 May;69(5):2638-50. doi: 10.1128/AEM.69.5.2638-2650.2003.
2
Cloning and thermostability of TaqI endonuclease isoschizomers from Thermus species SM32 and Thermus filiformis Tok6A1.嗜热栖热放线菌SM32和丝状栖热放线菌Tok6A1中TaqI核酸内切酶同裂酶的克隆及热稳定性
Biochem J. 1998 Jul 15;333 ( Pt 2)(Pt 2):425-31. doi: 10.1042/bj3330425.
3
Overproduction and crystallization of FokI restriction endonuclease.
FokI限制性内切酶的过量产生与结晶
Nucleic Acids Res. 1989 Nov 11;17(21):8741-53. doi: 10.1093/nar/17.21.8741.