Identification and Prokaryotic Expression of Anthocyanidin Synthase Gene in Prunus salicina
LV Shiheng
Abstract
LV Shiheng
Abstract
A cDNA encoded anthocyanidin synthase(ANS)was separated from a normalization and full-length cDNA library prepared previously with the ripe fruit of 'oil Lai'(Prunus salicina)by gene special primers designed from ANS genes of other Rosaceae plants.The cDNA named PsANS was 1 478 bp in full length and encoded a protein with 358 amino acids.The deduced amino acid sequence shares 98%,93% and 89% identity with those ANS from sweet cherry,apple and strawberry,respectively.Prokaryotic expression was conducted with the constructed vectors used PsANS and prokaryotic expression vector(pGEX-6P-1 and pET-32a)in E.coli BL21.The obtained fusion protein had a molecular weight consistent with that expected,and it further confirmed that the PsANS obtained by this experiment encoded ANS of 'oil Lai'(Prunus salicina).
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A cDNA encoded anthocyanidin synthase(ANS)was separated from a normalization and full-length cDNA library prepared previously with the ripe fruit of 'oil Lai'(Prunus salicina)by gene special primers designed from ANS genes of other Rosaceae plants.The cDNA named PsANS was 1 478 bp in full length and encoded a protein with 358 amino acids.The deduced amino acid sequence shares 98%,93% and 89% identity with those ANS from sweet cherry,apple and strawberry,respectively.Prokaryotic expression was conducted with the constructed vectors used PsANS and prokaryotic expression vector(pGEX-6P-1 and pET-32a)in E.coli BL21.The obtained fusion protein had a molecular weight consistent with that expected,and it further confirmed that the PsANS obtained by this experiment encoded ANS of 'oil Lai'(Prunus salicina).
Key concepts: Prunus salicina, Complementary DNA, Biology, Prunus, Gene, Anthocyanidin, Homology (biology), Molecular biology