Improvement of a PCRbased Method for Vector Construction of Multisitedirected Mutations in GC-rich Promoter

Journal of Capital Medical University ›› 2010, Vol. 31 ›› Issue (1): 88-92.

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Improvement of a PCRbased Method for Vector Construction of Multisitedirected Mutations in GC-rich Promoter

JIANG Luan¹, SHAO Lei², HU Yang³, DONG Ling-yue⁴

1. Class No. 3 of Grade 2005 in Department of 7Year Clinical Medicine, Capital Medical University; 2. Class No. 3 of Grade 2005 in Department of 5Year Clinical Medicine, Capital Medical University; 3. Class No. 4 of Grade 2005 in 5 Year's Clinical Medicine, Capital Medical University; 4. Department of Cell Biology, Basic Medical Sciences, School of Basic Medical Sciences, Capital Medical University

Received:1900-01-01 Revised:1900-01-01 Online:2010-02-21 Published:2010-02-21
Contact: DONG Ling-yue

Abstract

Abstract:

Objective To construct several vectors in which multi-site mutation occurs within a GC-rich hHSS promoter. Methods An interested genomic region of human hepatic stimulator substance(hHSS) was multi-site-directly mutated and amplified. During amplification, the melting temperature was reduced by adding certain concentration of DMSO and the concentration of the enzyme-digested products was increased through ethanol precipitation. Results The results showed that five different hHSS promoter vectors containing multipoint mutations were effectively constructed and confirmed by DNA sequencing. Conclusion This improved method for PCR amplification of GC-rich promoter was efficient, which provides a rapid, simple and economic approach to analysis of genomic DNA.

Key words: site directed mutagenesis, polymerase chain reaction, dimethyl sulfoxide, human hepatic stimulator substance

CLC Number:

Q 782

JIANG Luan;SHAO Lei;HU Yang;DONG Ling-yue. Improvement of a PCRbased Method for Vector Construction of Multisitedirected Mutations in GC-rich Promoter[J]. Journal of Capital Medical University, 2010, 31(1): 88-92.

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Improvement of a PCRbased Method for Vector Construction of Multisitedirected Mutations in GC-rich Promoter

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