Cloning and Prokaryotic Expression of Almond AcDHN1 Gene

【Objective】 The dehydrin gene was cloned and analyzed in order to identify the role of AcDHN1 gene in plant resistance of Amygdalus communis. 【Method】 Using Xinjiang cultivated almond 'zhipi' leaves, according to the reported DHN gene primers through the PCR technique to clone and analyze its prokaryotic expression of the AcDHN gene. 【Result】 we cloned a AcDHN1 (GeneBank accession NO.KT949395), this gene was 924 bp and encoding 308 amino acids, was a stable hydrophilic protein, belonging to Y₂K_n Dehydrin, speculated with 32.4 kD molecular weight, which subcellular localization was in nucleus. We connect the gene to prokaryotic expression vector to construct recombinant plasmid pET-AcDHN1, then we transformed the pET-AcDHN1 into E. coli BL21 (DE3) and induced. The SDS-PAGE result showed the protein was 52.7 kD in molecular weight and was consistent with the expected. 【Conclusion】 Through optimized the recombinant protein induction conditions we got the expressing condition was 0.1 mmol/L IPTG and 3 hours induction.