Cloning of Human Amelogenin Gene Encoding Mature Protein

Objective: The purpose of this study was to clone human amelogenin encoding mature protein from dental germ, that made step for expressing the recombinant human amelogenin in Escherichia coli. in the future. Methods: In the study, total RNA was extracred from the dental germ of a legally aborted embryo by Trizol, and synthesis of cDNA was obtained from the total RNA by RT-PCR with the Oliga primer, then the desired DNA products were conducted with PCR from cDNA .The segment (about 540bp) was inserted into expression vector PQE30 and the interesting plasmid was transformed into E.Coli. host DH5¦Á. 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 told that the human amelogenin gene encoding mature protein was inserted into vector PQE30 accurately. Conclusions: The human amelogenin encoding mature protein was conducted from dental germ of a legally aborted embryo. We got the recombinant plasmid which may express amelogenin gene for further research.