Subgroup A or B avian leukosis virus gp85 gene recombinant prokaryotic expression protein and purification method thereof, and applications

Abstract

The invention discloses a subgroup A or B avian leukosis virus gp85 gene recombinant prokaryotic expression protein and a purification method thereof, and applications. The method includes the following steps: (1) designing and synthetizing specific primers for amplifying gp85 genes; (2) obtaining gp85 gene fragments through PCR amplification, and performing purification; (3) separately cloning the gp85 genes of an ALV-A / B to pET-32a prokaryotic expression vectors, and performing sequencing and identification after screening positive clones; (4) recombining the expression of expression vectorpET-32a-A / B-gp85; (5) purifying expressed gp85 proteins through a urea method; and (6) recombining the renaturation of the gp85 proteins. Thus, soluble gp85 proteins with high purity can be obtained by using the method; and the expressed proteins can be taken as envelope antigens to establish an antibody detection method and used for preparing antiserum for immunizing animals, and also can provideraw materials for immunofluorescence, immunohistochemistry, etc.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a recombinant prokaryotic expression protein of the gp85 gene of subgroup A or B avian leukosis virus and its purification method and application. Background technique [0002] Avian leukemia (AL) is a variety of benign and malignant neoplastic diseases of poultry caused by avian leukosis virus (ALV) sarcoma virus group virus. Clinically, it is mainly characterized by immunosuppression, growth inhibition and tumors in multiple organs and tissues. According to host range, envelope protein antigenicity and cross-neutralization reaction, it was divided into 11 subgroups A-K, among which A-D, J and K subgroups were exogenous ALV. C and D subgroups are rare in the wild, E-I subgroup is endogenous ALV, and F-I subgroup mainly infects wild birds. Among the exogenous ALV, A, B and J subgroups have obvious pathogenicity to chickens, and are also more harmful to poultry production. Due to th...

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Problems solved by technology

Currently there is an ELISA antibody detection kit for ALV-A / B, but it is only used to detect the antibody level of ALV-A / B in chicken flocks, and cannot determine whether there is still ALV-A / B infection in chicken flocks

Serological monitoring in my country usually uses ELISA detection kits from companies such as the United States, which are expensive and limited in scope of promotion

The detection methods and kits developed according to foreign strains cannot accurately detect the infection status of domestic chickens, so the establishment of an applicable ALV-A / B detection system is particularly important for ALV purification.

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