Antibody Preparation of MsWRKY33 Protein and Response to Abiotic Stress

doi:10.7668/hbnxb.20192138

Abstract

Abstract: To explore the biological function of alfalfa MsWRKY33 in stress tolerance at the protein level,by using Medicago sativa cv.Zhongmu No.1 as the material,MsWRKY33 antigen sequence was cloned and its properties and functions were analyzed.The structural characteristics analysis and immunogenicity prediction of MsWRKY33 protein were performed by using bioinformatics.The structure and characteristics of MsWRKY33 protein analysis results showed that the full length of MsWRKY33 protein was 511 aa,which was mainly composed of random coil and extended chain structure.MsWRKY33 protein had no signal peptide and was a hydrophilic protein.The immunogenicity prediction results indicated that the protein was highly immunogenic.According to the specificity of the binding sequence,the appropriate segment 247-457 aa was selected,and the MsWRKY33 polyclonal antibody was prepared by using this segment through protein purification,immunization and other steps.Subsequently,Western Blotting method was used to analyze the expression of MsWRKY33 protein under four abiotic stress treatments:salt,cold,PEG and ABA,all could enhance the MsWRKY33 protein expression.Among them,NaCl brought the highest expression level,indicated that MsWRKY33 might play an important role in salt stress regulation.The test results provided a reference for further exploration of the function of MsWRKY33 and the mechanism regulating the stress tolerance of alfalfa.

Key words: Medicago sativa L., MsWRKY33, Polyclonal antibody, Western Blotting, Salt stress

CLC Number:

S541+.9
Q78

ZHANG Jinjin, WANG Yunping, WANG Xiao, ZHANG Shuxing, WANG Xuemin. Antibody Preparation of MsWRKY33 Protein and Response to Abiotic Stress[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(6): 63-70. doi: 10.7668/hbnxb.20192138.

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