Cloning and sequencing analysis of human PKC?Ⅱgene ORF
- VernacularTitle:人蛋白激酶C?Ⅱ基因开放读码框的克隆及序列分析
- Author:
Lian DUAN
- Publication Type:Journal Article
- Keywords:
PKC? gene;
PCR;
T/A cloning;
Sequence analysis
- From:
Journal of Chongqing Medical University
1987;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To clone human PRKCB1 gene open reading frame(ORF) in order to further research on it's function.Methods:RT-nested PCR method was adopted to the amplify the total length of human PRKCB1 ORF from human umbilical vein endothelial cell(HUVEC).T vector was inserted into the harvested fragment after a tail was added.With blue white screening,the gene ORF encoding human PKC?Ⅱ was obtained by special primers amplifying recombinant plasmids,and linked into T vector.Then the plasmid was identified by sequencing.Results:The human PRKCB1 ORF was amplified successfully with RT-nested PCR and T/A cloning,and the gene sequence was completely consistent with that reported in GenBank.Conclusion:Human PRKCB1 gene ORF was successfully cloned.The strategy of cloning may provide technical references for some genes hard to be cloned.
