关键词: 新疆红肉苹果')">新疆红肉苹果, MsMYB10转录因子, 序列分析, 原核表达

Abstract:

【Objective】 Cloning, sequence analysis and expression in E. coli of MsMYB10 gene from Malus sieversii f. neidzwetzkyana was conducted to further explore the red development mechanism and breed cultivars of red-flesh apple. 【Method】A cDNA fragment about 700 bp was amplified from the total RNA of leaves of Malus sieversii f. neidzwetzkyana by reverse transcription PCR(RT-PCR) with a pair of specific primers based on the sequences of MdMYB10. The recombinant prokaryotic expression vector pET30a-MsMYB10 was constructed by inserting the cDNA fragment into the prokaryotic expression vector pET-30a, and then transformed into E. coli BL21(DE3). 【Results】 Sequence analysis showed that the fragment contains a full coding region of 732 bp encoding 244 amino acid residues with a molecular mass of 28.56 kD. Its GenBank accession number is GQ500894. This deduced protein has a pI of 8.41. Results of the study showed that MsMYB10 exhibited typical features of the R2R3-MYB domain. Tryptophan residues are conserved in R2R3 domain. There is one transcriptional activation domain rich in acidic amino acids in the C-terminal. MsMYB10 exhibits a homology of 99% and 98% with MdMYB10 in nucleotide and amino acid levels, respectively. MsMYB10 has no signal peptide, but a nuclear localization signal. The homology tree showed that MsMYB10 is at the same evolutionary branch with MdMYB10. The SDS-PAGE displays that the expressed proteins consistent with the size of expected protein. 【Conclusion】 MsMYB10 was cloned from Malus and expressed in E. coli. These results have provided a foundation for further purifying and identifying target protein and function study of MsMYB10.

Key words: Malus sieversii f. neidzwetzkyana')">Malus sieversii f. neidzwetzkyana, MsMYB10, sequence analysis, prokaryotic expression