大豆查尔酮异构酶基因的克隆及乳酸菌表达载体的构建

Liu Hongyu, Biotechnological Center of Jilin Agricultural University, Changchun; Wang Piwu, Biotechnological Center of Jilin Agricultural University, Changchun; Fu Yongping, Biotechnological Center of Jilin Agricultural University, Changchun

Cloning of tap Ⅱchalcone isomerases (CHI1 A) gene and construction of Lactococcus Lactis expression vector | 大豆查尔酮异构酶基因的克隆及乳酸菌表达载体的构建

2010

Liu Hongyu, Biotechnological Center of Jilin Agricultural University, Changchun (China) | Wang Piwu, Biotechnological Center of Jilin Agricultural University, Changchun (China) | Fu Yongping, Biotechnological Center of Jilin Agricultural University, Changchun (China)

[Objective] The aim was to clone Tap Ⅱ chalcone isomerases (CHI1 A) from soybean specially and construct expression vector of PNZ8149- CHI1 A , and then transform it into Lactococcus Lactis NICE systers. [Method] Chalcone isomerases (CHI1 A) was cloned by RT-PCR method, and it was sequenced after cloning into pMD18-T vectors, and recombined to expression vector PNZ8149- CHI1 A, then it was transformed into Lactococcus Lactis NZ3900 [Result] The sequencing results indicated that the cloned fragment of CHI1 A contained 670 nucleotides, and shared a sequence homology of 92% with that from Genbank accession number AF595413 (CHI1 A) . CHI1 A was transformed into NICE expression system successfully by identification of PCR and digestion. [Conclusion] The foundation of using the microorganism fermentation method to produce flavonoids was laid by construction of efficient induction expression vector with chalcone isomerases CHI1 A.

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