Cloning of ACC Oxidase Gene of Carnation and Construction of Its Plant Expression Vectors
Yixun Yu, Junwei Zhang, Zhenyuan Sun, Manzhu Bao
Abstract
Yixun Yu, Junwei Zhang, Zhenyuan Sun, Manzhu Bao
Abstract
A pair of primers were designed according to the 1 aminocyclopropone 1 carboxylic acid (ACC) oxidase gene of carnation and the primers were used to amplify the genomic DNA fragment of about 1.2 kb by polymerase chain reaction (PCR)by taking genome DNA from 'American' carnation leaves as template. The PCR product was cloned into T tailing pMD18 vector. Sequencing indicated that the ACC oxidase gene included three exons interrupted by 2 introns with identical positions as they are in tomato. ACC oxidase gene were respectively cloned into plant expression vector pMOGMON in sense and antisense orientation. Recombinant expression vectors were identified by restriction enzyme and PCR analysis. PCR indicated plant expression vectors were transferred into A. tumefaciens .
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A pair of primers were designed according to the 1 aminocyclopropone 1 carboxylic acid (ACC) oxidase gene of carnation and the primers were used to amplify the genomic DNA fragment of about 1.2 kb by polymerase chain reaction (PCR)by taking genome DNA from 'American' carnation leaves as template. The PCR product was cloned into T tailing pMD18 vector. Sequencing indicated that the ACC oxidase gene included three exons interrupted by 2 introns with identical positions as they are in tomato. ACC oxidase gene were respectively cloned into plant expression vector pMOGMON in sense and antisense orientation. Recombinant expression vectors were identified by restriction enzyme and PCR analysis. PCR indicated plant expression vectors were transferred into A. tumefaciens .
Key concepts: Biology, Carnation, Gene, Molecular biology, Cloning (programming), Expression vector, Restriction enzyme, Complementary DNA