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- W2385936701 abstract "Objective:To construct recombinant expression plasmids of cDNAs encoding human protein kinase CK2α,α′and β subunits for further investigation on the structure and function of CK2 Methods:RT PCR,directed cloning and DNA sequencing were utilized Results:The cDNAs encoding human protein kinase CK2 α,α′and β subunits were obtained from human promyelocytic leukemia cells (HL 60)by RT PCR,Nde I /Hind Ⅲ digested PCR products were then ligated to the expression vector pT7 7 Transformants were taken from E coli DH5α using CaCl 2 method and then screened with gel electrophoresis The results of restriction enzyme digestion analysis indicated that the sizes of 3 insert fragments and 3 recombinant plasmids were consistent with the predicated values Four positive clones of each CK2 subunit recombinant were randomly selected,purified by polyethylene glycol method and sequenced with various primers in two directions by way of Big Dye TM terminator cycle sequencing ready reaction kit Two,one and two of four clones in each CK2 subunit recombinant possessed correct sequences of coding region of human protein kinase CK2α,α′and β subunits,which were termed as pTCKA,pTCKA′and pTCKB,respectively;while the others contained one or two base mutants Conclusion:It is suggested that successful cloning of pTCKA,pTCKA' and pTCKB lays a solid basis on the study of the expression of human protein kinase CK2α,α′and β subunits in prokaryotes and their possible uses as probes" @default.
- W2385936701 created "2016-06-24" @default.
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- W2385936701 date "2002-01-01" @default.
- W2385936701 modified "2023-09-25" @default.
- W2385936701 title "Cloning and sequencing of recombinant cDNAs encoding 3 subunits of human protein kinase CK2" @default.
- W2385936701 hasPublicationYear "2002" @default.
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