[Expression of protein kinase C alpha in Chinese hamster ovary cells].
OBJECTIVE
To study the expression of protein kinase C (PKC) alpha subtype in Chinese hamster ovary (CHO) cells.


METHODS
The PKC alpha primer pairs were designed based on the GenBank sequence of PKC alpha of a species with the highest homology to Chinese hamster identified using EMBL Data Library Clustalw tool. The sequence coding for PKC alpha, amplified from the CHO cells using RT-PCR, was ligated to the pGEM-T plasmid vector, and the recombinant vector was transformed into E.coli DH5alpha with the positive colones selected by blue/white screening. Restriction enzyme digestion, gel electrophoresis analysis, followed by sequencing of the digestion products were performed for identification of the recombinant. Western blotting was used to analyze the PKC alpha expression in the CHO cells.


RESULTS
The presence of PKC alpha mRNA was detected in the CHO cells by RT-PCR. Western blotting also identified PKC alpha expression in the cells.


CONCLUSIONS
PKC alpha expression has been identified in the CHO cells, which may facilitate further structural and functional study of PKC alpha and investigation of its role in the intracellular signal transduction pathways.
Dong
Li-jin
LJ
Department of Pharmacology, Medical College of Chinese People's Armed Police Force, Tianjin 300162, China. dongdonglijin@126.com
Zhang
Hai-lin
HL
Li
Chu-yan
CY
D000818:Animals; D015153:Blotting, Western; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D051571:Protein Kinase C-alpha / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D051571:Protein Kinase C-alpha