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Published on: July 10, 2014
[Cloning of human amelogenin gene encoding mature peptide]
Lan Cheng1, Jian-qiang Lei, Qi-quan Zhu
1Department of Operation Dentistry,School of Stomatology,Ninth People's Hospital, Shanghai Second Medical University, Shanghai 200011, China.
Purpose:
The purpose of this study was to clone human amelogenin gene encoding mature protein, which provides a basis for expressing the recombinant human amelogenin in Escherichia coli. in the future.
Methods:
In this study, total RNA was extracted from the dental germ of a legally aborted embryo by Trizol. Using RT-PCR technique we obtained synthesis of cDNA from the total RNA, and the desired DNA products were conducted with PCR from cDNA. The segment was inserted into expression vector PQE30 and the interesting plasmid was transformed into Escherichia coli. host DH5alpha. The double-stranded DNA of positive clone was analyzed by PCR, restriction endonuclease mapping and DNA sequence analysis.
Results:
The sequence analysis of recombinant plasmid showed that the human amelogenin encoding mature protein was inserted into vector PQE30 accurately.
Conclusions:
We conducted human amelogenin encoding mature protein from dental germ of a legally aborted embryo and got the recombinant plasmid which may express amelogenin gene for further research.
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