Construction and identification of the fusion clone of the SCF 5’flanking sequence and its full-length cDNA
TAN Yun-nian
TAN Wen-bin
PENG Xing-hua
Abstract：Objective：To study the effect of different length DNA sequences of the 5’SCF 1．42 kb flanking sequence in the expression and regulation of full-length cDNA in eukaryotic cells，a fused clone of the 5’SCF 1．42kb flanking sequence and its full-length cDNA were constructed．Methods：A 1．42 kb flanking sequence and a 1．2 kb full-length cDNA were achieved by PCR from human genomic DNA and by RT-PCR from HepG2 mPNA，respectively，and then cloned into pGEM-T cloning vector and identified．Both clones were digested with fitly restricted endonuclease and three DNA fragments，480 bp，980 bp，1．2 kb cDNA were harvested．Finally，these three DNA fragments were subcloned into pUC19 cloning vector in turn．Results：The fused clone of 5’SCF 1．42 kb flanking sequence and its full-length cDNA were successfully constructed．Conclusion：To avoid the interruption of some restricted endonuclease sites，the way to cut larger fragments into smaller fragments is still useful and effective in the process of gene cloning
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Publication Date：2001-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 6-9 )