ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2011, Vol. 42 ›› Issue (9): 1302-1308.doi:

• 预防兽医 • Previous Articles     Next Articles

Prokaryotic Expression of Tm7 Gene of Taenia multiceps and Establishment of Indirect ELISA Using the Expressed Protein

AN Xiao-xue1, YANG Guang-you1*, WANG Ying-wang1, MU Jing1,

YANG Ai-guo3, GU Xiao-bin1, YANG Ying-dong2, WEI Lei-fei2,

WEN Jianguo2, WANG Shuxian1, BIAN Yao 4

  

  1. 1.College of Veterinary Medicine,Sichuan Agricultural University, Ya’an 625014, China；2.Panzhihua Animal Science and Technology Institute, Panzhihua 617061, China；3. Preventive and Control Center for Animal Disease of Sichuan Province, Chengdu 610041, China； 4. Yucheng District Animal Husbandry Bureau of Sichuan Province, Ya’an 625000, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2011-09-23 Published:2011-09-23
  • Contact: YANG Guang-you

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Abstract: In order to establish diagnostic method of coenuriasis，total RNA was extracted from protoscoleces of cyst which were recovered from the brain of sheep. The complete sequence of Tm7 was amplified and sequenced. The open reading frame of Tm7 was then amplified, its ORF consisting of 207 bp and encoding 68 amino acids. A recombinant plasmid pET-32a-Tm7 was constructed and transformed into E. coli BL 21 for in vitro expression. SDSPAGE and Western blot were employed for analyzing the recombinant protein. The purified recombinant protein was used for development of an indirect ELISA for detection of anticoenuriasis antibodies. The results showed that target fusion protein was expressed in E. coli, it was about 27 kD in molecular weight and could be recognized by coenurosis positive serum. Futhermore, in indirect ELISA this recombinant protein showed 93.3% of sensitivity and 94.1% of sensitivity, indicated it can be used as diagnostic antigen.