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Construction of prokaryotic expression vector and expression of GbMYBFL protein

By: Liangqiong Ma, Weiwei Zhang, Feng Xu, Yongling Liao

Key Words: Ginkgo biloba, GbMYBFL, prokaryotic expression, PET32a, SDS-PAGE.

Int. J. Biosci. 13(2), 104-109, August 2018.

DOI: http://dx.doi.org/10.12692/ijb/13.2.104-109

Certification: ijb 2018 0166 [Generate Certificate]

Abstract

Flavonoids are one of the main secondary metabolites of Ginkgo biloba, and MYB is a transcription factor involved in regulation of flavonoid synthesis. In order to construct the prokaryotic expression vector of GbMYBFL from Ginkgo biloba and express the target protein, a pair of primers were designed with Bam H I and EcoR I restriction sites to amplify the ORF region of Ginkgo GbMYBFL gene by PCR. The PCR product was digested by BamH I and EcoR I, and inserted into the prokaryotic expression vector pET32a. Then the recombinant plasmid was transformed into E.coli BL21 (DE3). After induction and expression, the protein was detected by SDS-PAGE. The results showed that the prokaryotic expression recombinant plasmid pET32a-GbMYBFL was successfully constructed. The recombinant plasmid was induced by IPTG in E. coli BL21 (DE3), and the size of the protein electrophoresis band is about 48 kDa, which was consistent with the molecular weight of protein predicted by bioinformatics tools. This study laid the foundation for the functional research of GbMYBFL in Ginkgo biloba.

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Construction of prokaryotic expression vector and expression of GbMYBFL protein

Aharoni A, De Vos C, Wein M, Sun Z, Greco R, Kroon A, Mol JM, O’Connell AP. 2001. The strawberry FaMYB1 transcription factor suppresses anthocyanin and flavonol accumulation in transgenic tobacco. The Plant Journal 28(3), 319-332.

https://doi.org/10.1046/j.1365-313X.2001.01154.x

Albert NW, Lewis DH, Zhang H, Schwinn KE, Jameson PE, Davies KM. 2011. Members of an R2R3-MYB transcription factor family in Petunia are developmentally and environmentally regulated to control complex floral and vegetative pigmentation patterning. The Plant Journal 65(5), 771-784.

https://doi.org/10.1111/j.1365-313X.2010.04465.x

Borevitz JO, Xia Y, Blount J, Dixon RA, Lamb C. 2000. Activation tagging identifies a conserved MYB regulator of phenylpropanoid biosynthesis. The Plant Cell Online 12(12), 2383-2393.

https://doi.org/10.1105/tpc.12.12.2383

Deluc L, Barrieu F, Marchive C, Lauvergeat V,  Decendit A, Richard T, Carde JP, Mérillon JM. 2006. Characterization of a grapevine R2R3-MYB transcription factor that regulates the phenylpropanoid pathway. Plant physiology 140(2), 499-511.

https://doi.org/10.1104/pp.105.067231

Deluc L, Bogs J, Walker AR, Ferrier T, Decendit A, Merillon JM, Robinson SP, Barrieu F. 2008. The transcription factor VvMYB5b contributes to the regulation of anthocyanin and proanthocyanidin biosynthesis in developing grape berries. Plant physiology 147(4), 2041-2053.

https://doi.org/10.1104/pp.108.118919

Espley RV, Hellens RP, Putterill J, Stevenson DE, Amma SK, Allan AC. 2007. Red colouration in apple fruit is due to the activity of the MYB transcription factor, MdMYB10. The Plant Journal 49(3), 414-427.

https://doi.org/10.1111/j.1365-313X.2006.02964.x

Feng S, Wang Y, Yang S, Xu F, Chen X. 2010. Anthocyanin biosynthesis in pears is regulated by a R2R3-MYB transcription factor PyMYB10. Planta 232(1), 245-255.

https://doi.org/10.1104/pp.108.118919

Martin C, Paz-Ares J. 1997. MYB transcription factors in plants. Trends in Genetics 13(2), 67-73.

https://doi.org/10.1016/S0168-9525(96)10049-4

Penfield S, Meissner RC, Shoue DA, Carpita NC, Bevan MW. 2001. MYB61 is required for mucilage deposition and extrusion in the Arabidopsis seed coat. The Plant Cell Online 13(12), 2777-2791.

https://doi.org/10.1105/tpc.010265

Qin Q. 2007. Molecular cloning and characterization of transcription factors involved in liginin biosynthetic pathway and phenylpropanoid pathway in Ginkgo biloba. PhD thesis, Fudan Univesity, 60-75.

Quattrocchio F, Wing J, van der Woude K, Souer K, de Vetten N, Mol J, Koes R. 1999. Molecular analysis of the anthocyanin2 gene of petunia and its role in the evolution of flower color. The Plant Cell Online 11(8), 1433-1444.

https://doi.org/10.1105/tpc.11.8.1433

Takos AM, Jaffé FW, Jacob SR, Bogs J, Robinson SP, Walker AR. 2006. Light-induced expression of a MYB gene regulates anthocyanin biosynthesis in red apples. Plant physiology 142(3), 1216-1232.

https://doi.org/10.1104/pp.106.088104

Urao T, Yamaguchi-Shinozaki K, Urao S, Shinozaki K. 1993. An Arabidopsis myb homolog is induced by dehydration stress and its gene product binds to the conserved MYB recognition sequence. The Plant Cell 5(11), 1529-1539.

https://doi.org/10.1105/tpc.5.11.1529

Wang ZY, Kenigsbuch D, Sun L, Harel E, Ong MS, Tobin EM. 1997. A Myb-related transcription factor is involved in the phytochrome regulation of an Arabidopsis Lhcb gene. The Plant Cell Online 9(4), 491-507.

https://doi.org/10.1105/tpc.9.4.491

Xu F, Ning Y, Zhang W, Liao Y, Li L, Cheng H, Cheng S. 2013. An R2R3-MYB transcription factor as a negative regulator of the flavonoid biosynthesis pathway in Ginkgo biloba. Functional & integrative genomics 14(1), 177-189.

https://doi.org/10.1007/s10142-013-0352-1

Yusuke B, Chikako H, Yoshimichi H, Megumi I, Hideo B, Takaya M. 2007. Isolation and functional analysis of a MYB transcription factor gene that is a key regulator for the development of red coloration in apple skin. Plant and Cell Physiology 48(7), 958-970.

https://doi.org/10.1093/pcp/pcm066

Zhang W, Xu F, Cheng S, Liao Y. 2018. Characterization and functional analysis of a MYB gene (GbMYBFL) related to flavonoid accumulation in Ginkgo biloba. Genes & genomics 40(1), 49-61.

https://doi.org/10.1007/s13258-017-0609-5

Liangqiong Ma, Weiwei Zhang, Feng Xu, Yongling Liao .
Construction of prokaryotic expression vector and expression of GbMYBFL protein.
Int. J. Biosci. 13(2), 104-109, August 2018.
https://innspub.net/ijb/construction-prokaryotic-expression-vector-expression-gbmybfl-protein/
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