A MutS-based protein chip for detection of DNA mutations

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	A MutS-based protein chip for detection of DNA mutations
	Bi, LJ ; Zhou, YF ; Zhang, XE ; Deng, JY ; Zhang, ZP ; Xie, B ; Zhang, CG
刊名	ANALYTICAL CHEMISTRY
	2003
卷号	75 期号:16 页码:4113-4119
ISSN号	0003-2700
通讯作者	Zhang, XE, Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China
中文摘要	This paper describes a new protein chip method for detection of single-base mismatches and unpaired bases of DNA, using a genetic fusion molecular system Trx-His(6)-Linker peptide-Strep-tagII-linker peptide-MutS (THLSLM). The THLSLM coding sequence was constructed by attaching Strep-tag 11 and mutS gene to pET32a (+) sequentially with insertion of a linker peptide coding sequence before and behind Strep-tagII gene, respectively. THLSLM was expressed in E. coli AD494 (DE3) and purified using Ni2+-chelation affinity resin. THLSLM retained both mismatch recognition activity and streptavidin binding affinity. THLSLM was then immobilized on the chip matrix coated with streptavidin through the Strep-tag II-streptavidin binding reaction. The resulting protein chip was used to detect the mismatched and unpaired mutations in the synthesized oligonucleotides, as well as a single-base mutation in rpoB gene from Mycobacterium tuberculosis, with high specificity. The method could potentially serve as a platform to develop the high-throughput technology for screening and analysis of genetic mutations.
学科主题	Chemistry, Analytical
收录类别	SCI
语种	英语
WOS记录号	WOS:000184798100020
公开日期	2011-09-23
内容类型	期刊论文
源URL	[http://210.72.129.5/handle/321005/55379]
专题	沈阳应用生态研究所_沈阳应用生态研究所_期刊论文
推荐引用方式 GB/T 7714	Bi, LJ,Zhou, YF,Zhang, XE,et al. A MutS-based protein chip for detection of DNA mutations[J]. ANALYTICAL CHEMISTRY,2003,75(16):4113-4119.
APA	Bi, LJ.,Zhou, YF.,Zhang, XE.,Deng, JY.,Zhang, ZP.,...&Zhang, CG.(2003).A MutS-based protein chip for detection of DNA mutations.ANALYTICAL CHEMISTRY,75(16),4113-4119.
MLA	Bi, LJ,et al."A MutS-based protein chip for detection of DNA mutations".ANALYTICAL CHEMISTRY 75.16(2003):4113-4119.