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Characterization and Mutation of Soybean MYB Transcription
Factor GmMYBZ2 and Its Prokaryotic Expression
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Abstract  Soybean (Glycine max) GmMYBZ2 is one of typical plant R2R3-MYB family genes. In this study, its protein characterization and transcriptional activity were analyzed by DNAMAN and yeast (Saccharomyces cerevisiae) one-hybrid system, respectively. Moreover, the prokaryotic expression vector was constructed and expressed in Escherichia coli as well. Results showed that GmMYBZ2 not only exhibited typical features of the R2R3-MYB transcription factors, but also had a conserved amino acid motif and a predicted zinc finger region at its C-termini. Genomic DNA sequence analysis showed that it contained an intron in the R3 domain which was located at 263~395 bp. In order to investigate the infections of these motifs in GmMYBZ2’s transcriptional activity, the conserved motif and zinc finger region were detected by PCR. The effect of the altered proteins were monitored using yeast one-hybridize, which showed that GmMYBZ2 had transcriptional activation function. And the intron, conserved motif and zinc finger region might repress the transcriptional activity of GmMYBZ2. Moreover, the encoding sequence of GmMYBZ2 gene was inserted into pET30a to construct prokaryotic expression vector pET-ZD2 which was then transformed into E. coli BL21 (DE3) pLysS and Rosetta2(DE3) pLysS, respectively. Results showed that GmMYBZ2 could be expressed in E. coli Rosetta2(DE3) which contained complementary rare codons successfully.
Key wordsMYB transcription factor      mutation      yeast one-hybrid      prokaryotic expression     
Received: 06 May 2008     
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http://journal05.magtech.org.cn/Jwk_ny/EN/      OR     http://journal05.magtech.org.cn/Jwk_ny/EN/Y2009/V17/I2/301
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