Cloning and PRokaRyotic ExpRession of ATG8 ( a MoleculaR MaRkeR of Autophagy) Gene fRom Wheat and PRepaRation of AntiseRum

doi:10.3969/j.issn.1000-7091.2013.01.019

Abstract

Abstract: Total RNA was isolated fRom wheat vaRiety L10 and ReveRse tRanscRiption by using the anchoRed pRimeRs in this papeR to fuRtheR undeRstand the biological function of the autophagy gene ATG8 fRom wheat． And then the ATG8 weRe amplified by PCR and then cloned into the PMD19-T vectoR，one of which was subcloned into the expRession vectoR． The Recombinant plasmid was identified by sequencing and digestion of RestRiction enzymes． The Recombinant expRession vectoR was constRucted and tRansfoRmed into E． coli stRain Roseta( DE3) subsequently， then thRough IPTG-induction in host bacteRia E． coli Rosetta-gami B( DE3) and detected by SDS-PAGE． The Rabbit anti- ATG8 antibody was pRepaRed and was detected by WesteRn Blotting analysis． The ATG8 weRe obtained paRtly and successfully expRessed in the pRokaRyotic expRession system． The expeRiment offeRed foundation to the studying of autophagy and function of ATG8 gene in wheat．

Key words: ATG8, WesteRn Blotting, Anti-seRum, Fusion pRotein, PRokaRyotic expRession

CLC Number:

Q942

WU Hong-bo, LIU Gang, ZHANG Lu-lu, WANG Dong-mei. Cloning and PRokaRyotic ExpRession of ATG8 ( a MoleculaR MaRkeR of Autophagy) Gene fRom Wheat and PRepaRation of AntiseRum[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2013, 28(1): 101-105. doi: 10.3969/j.issn.1000-7091.2013.01.019.

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