Construction of prokaryotic expression clone for pig amelogenin gene encoding mature peptide

doi:10.3877/cma.j.issn.1674-1366.2008-02-003

Abstract

Abstract:

Objective

To clone pig amelogenin gene encoding mature peptide， and construct prokaryotic expression clone which lay the foundation for expressing the recombinant pig amelogenin in Escherichia coli. in the future.

Methods

Total RNA was extracted from the dental germ of a nascent pig by Trizol. The cDNA fragment of pig Amelogenin gene was obtained with RT-PCR from total RNA. The segment was inserted into prokaryotic gene fusion vector PGEX4T1 and the interesting plasmid was transformed into Escherichia coli. host DH5α. The double-stranded DNA of positive clone was characterized by restriction endonuclease mapping and DNA sequence analysis.

Results

The sequence analysis of recombinant plasmid showed that the pig amelogenin encoding mature protein was inserted into vector PGEX4T1 accurately.

Conclusions

The recombinant prokaryotic expression plasmid PGEX4T1-pAm was successfully constructed by properly inserting pig amelogenin gene encoding mature peptide.

Key words: Amelogenin, RT-PCR, Recombinant plasmid

Lan CHENG, Rong SHU. Construction of prokaryotic expression clone for pig amelogenin gene encoding mature peptide[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2008, 02(02): 116-121.

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Figures/Tables 3

References 13

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