Scientia Agricultura Sinica ›› 2007, Vol. 40 ›› Issue (9): 2084-2090 .

• ANIMAL SCIENCE·VETERINARY SCIENCERE·SOURCE INSECT • Previous Articles     Next Articles

Expression and Purification of Goose’s HSP70 and Complex Formation with Virus Polypeptide

  

  1. 青岛农业大学优质水禽研究所
  • Received:2006-05-11 Revised:2006-10-24 Online:2007-09-10 Published:2007-09-10

Abstract: Abstract:【OBJECTIVE】 In order to study the specificity expression of HSP70 in heat shock, HSP70 purification and its characteristic of coalescent with antigen peptide to form the compound, we designed this experiment;【METHOD】 60 healthy, 6 weeks ages male Wulong geese were selected and stochastically divided into 3 groups, the geese in Control group didn’t pass through the heat treatment, while the treatment 1 group were shocked by 1 time 42±1℃, 5h acute heat treatment and then were slaughtered immediately, the treatment 2 group restored 12h after the heat treatment under the normal raising condition and then were slaughtered. Hearts were taken to make paraffin section for the immunization organizations chemical experiment and livers were used to purify HSP70. Geese hearts HSP70 expression differences in 3 groups were determined, at the same time experiments of HSP70 purification and appraisal in the liver was carried on. HSP70 purification and synthesis HBV PreS1 multi- peptides unified the compound, and was determined by the double special immune body ELISA. 【RESULTS】 The findings indicated that, widespread HSP70 masculine pellets in the cardic muscle were found in the hot shock condition, HSP70 expression in treatment group 1 centered in the karyotheca and its periphery, but treatment group 2 centered the emphasis in the cell membrane around; We could obtain the HSP70 purification through two sets of purification plans; the synthesis peptide and the HSP70 purification forms the compound under the certain condition, examination taken to compound formation with the double specificity immune body, and the examination result was masculine, which explained that the compound had formed. 【CONCLUSION】 HSP70 specificity expression in heat shock shifted by time, and it may suggest that HSP70 possibly do some certain functions in the cell protection; we can obtain high-purity HSP70 protein under the low pressure chromatographic analysis condition, and it was compared to be better the molecular sieve preliminary separation, ConA-agarose chromatographic analysis and the ADP-agarose chromatographically analyzes flow; Under in vitro certain condition, the man-power synthesizes the peptide to be possible to unify with HSP70 forms the compound, this for further proved the compound the immunity function provides the experimental foundation.

Key words: Heat shock, goose, HSP70, expression, purification, Virus complexs

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