Method for preparing swine fever recombinant subunit vaccine

• Main Authors: TIAN HONG, SHANG YOUJUN, YIN SHUANGHUI, LIU YANHONG, HAN XUEQING, LIU XIANGTAO, SUN SHIQI
• Format: Patent
• Language: Chinese; English
• Published: 06.04.2011
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; HUMAN NECESSITIES; HYGIENE; MEDICAL OR VETERINARY SCIENCE; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS ORMEDICINAL PREPARATIONS; SPIRITS; VINEGAR; WINE

The invention relates to a method for preparing a recombining subunit vaccine of swine fever. The method comprises the following steps: A. a reverse transcription-polymerase chain type reaction method is utilized to clone a coding gene of vaccine strain E2 protein of the swine fever; B. a E2 gene is constructed and fused; C. a recombining expression vector rE2 is constructed; D. rE2 induction expression is formed; E. the rE2 is purified; and F. the activity of the rE2 is detected through a Western blotting method. The method creatively uses a double-E2 gene serial expression mode in order that the number of E2 epitope is doubled, thereby remarkably improving the immune effect of the recombining E2 subunit vaccine; to improve the expression efficiency of double E2 protein, a colibacillus RBS sequence (5'-AAGGAG-3') is added on the upstream of an initial codon (ATG) of 5'end of the second E2 gene; at the same time, an amino acid joint 5'-GSA GSA AGS GEF-3' is added between E2 genes in serial connection so that