Construction of a middle fragment-deleted class II molecule transactivator mutant by modified OE-PCR technique
- Author:
Zheng GUAN
1
Author Information
1. Department of Clinical Diagnosis
- Publication Type:Journal Article
- From:
Academic Journal of Second Military Medical University
2006;27(9):1014-1017
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To develop a simple and efficient method for constructing a middle fragment-deleted mutant of MHC class II molecule transactivator (C II TA) mutant with the 109th to 226th amino acid codons deleted. Methods: Two gene fragments at each end of the deleted C II TA gene were obtained by OE-PCR method and were mixed together for 8 PCR cycles without primers to achieve effective overlapping, then 2 primers was added for amplification of the desired fragments. The amplification products were subsequently cloned into eukaryotic vector pIRES for identification. Results: A mutant of C II TA with the 109th to 226th amino acids deleted was successfully constructed. Conclusion: This modified OE-PCR technique overcomes some shortcomings of traditional method and is very suitable for constructing mutants with middle fragment deletion, making it worth to be popularized.
