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- W2347830391 abstract "Objective: In order to obtain a high level express of human IL 3,we modified the human IL 3 cDNA with PCR based mutagenesis method. And also we have determined the new mutant variant human IL 3 cDNA structure and its protein products activity. This method can be used to study gene modification, the structure or function of protein and the relationship between themselves.Methods: The human IL 3 cDNA mutants were obtained by PCR in vitro. The codon Met3 and Lys116 in human IL 3 cDNA were substituted by codon (GTT) of Val. Nucleotide sequences of the PCR products were determined by DNA sequence.Natural human IL 3 cDNA (NhIL 3 cDNA) and mutant variant human IL 3 cDNA (MvhIL 3 cDNA) have been inserted downstream of promoter with a set of expression vectors, and transfered into E.Coli.Results: E.Coli bacterial cells boring the plasmid pSM53 NhIL 3 expressed only 7% protein of the total bacterial body protein but the pSM53 MhIL 3 cDNA had produced a high level of human IL 3 about 16% of the total bacterial body protein. The molecular weight of the product is 15 kD. Their biological activity respectively is 1.5×10 4 U/ml and 6.9×10 4 U/ml。Conclusion:The human IL 3 cDNA can be reconstucted with PCR mutagenesis method to get a new human IL 3 cDNA mutant. The activity of this MVhIL 3 expressed protein we have obtained is 3 4 times of the NhIL 3." @default.
- W2347830391 created "2016-06-24" @default.
- W2347830391 creator A5041971287 @default.
- W2347830391 date "1999-01-01" @default.
- W2347830391 modified "2023-09-25" @default.
- W2347830391 title "PCR site directed mutagenesis of human IL 3 cDNA in vitro human IL 3 by PCR site directed mutagenesis study" @default.
- W2347830391 hasPublicationYear "1999" @default.
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