In order to identify host factors which interact with the movement protein (MP) of Apple chlorotic leaf spot virus (ACLSV), ACLSV MP was cloned into the bait vector pGBKT7 and used to screen a cDNA library of Malus sylvestris cv. R12740-7A, which had previously been constructed by yeast two-hybrid sequencing transformation. The protein functions of the identified host factors were determined according to their gene annotations in GenBank. The result showed that the bait plasmid pGBKT7-MP showed no virulence or self-activating effect on yeast strain Y2H Gold. Sixty-nine interactor proteins were identified, which were divided into the following 10 classes according to their described functions: hydrolases; pathogenesis-related proteins; DNA binding proteins; phosphatases; ligases; proteins with catalytic activity; phenylalanine ammonialyases; peroxidases; NAD binding proteins; and proteins of unknown function. Bioinformatic analysis of gene homology suggested that phosphatases, pathogenesis-related proteins and glyceraldehyde-3-phosphate dehydrogenase A may play an important role in the interaction between virus and host. This study may provide a theoretical basis for the further study of viral pathogenesis and virus-host interaction mechanisms.
Flexiviridae ; genetics ; metabolism ; Malus ; genetics ; metabolism ; virology ; Molecular Sequence Data ; Plant Diseases ; genetics ; virology ; Plant Proteins ; genetics ; metabolism ; Plant Viral Movement Proteins ; genetics ; metabolism ; Protein Binding ; Two-Hybrid System Techniques

Objective To observe the effect of Ningxia aqueous extracts of fruitless lycium sprout (AEFLS) on cardiomyo cyte antioxidation and apoptosis-related protein expression in aging mice.Methods The natural aging C57BL/6J mice with 13 months old were randomly divided into aged control group,AEFLS low dose group (AEFLS1),AEFLS middle dose group (AEFLS2)and AEFLS high dose group(AEFLS3).The AEFLS1,AEFLS2 and AEFLS3 groups were respectively given with 5,10,20 mg/kg AEFLS gavage,while the aged control group was given with the normal saline gavage,for continuous8 weeks.The xan thine oxidase assay and thiobarbituric acid method were used for the determination of SOD and MDA in heart tissues.Western-blot and immunohistochemical method were used to detect the expressions of Bcl-2,Bax and Capase-3 in heart tissue.Results Compared with the aged control group,the MDA level in the AEFLS2 and AEFLS3 groups was decreased,while the SOD activity was increased,the difference was statistically significant (P＜0.01).The Western-blot result showed that compared with the aged control group,the optical density value of Bcl-2 in the AEFLS2 and AEFLS3 groups was increased,but the optical density values of Bax and Capase-3 were decreased (P＜0.01);the immunohistochemical results showed that compared with the aged control group,the immunopositive(IP) expressions of Bcl-2 protein in heart tissues in the AEFLS2 and AEFLS3 groups were increased (P＜0.01),while the IP expressions of Bax and Capase-3 were decreased (P＜0.01).Conclusion Middle and high doses of AEFLS can increase the antioxidative ability of myocardial tissue,up-regulates the Bcl-2 expression,down-regulates the Bax and Capase-3 expressions and plays anti-cardiomyocyte apoptotic role.