TY  - JOUR
T1  - Expression of Recombinant TB10.4 of <I>Mycobacterium bovis</I> and the Preliminary Study of its Potential in Diagnosis of Bovine Tuberculosis
AU - Liu, Shuqing AU - Gao, Xintao AU - Li, Pingjun AU - Jia, Hong AU - Xin, Ting AU - Hou, Shaohua AU - Guo, Xiaoyu AU - Yuan, Weifeng AU - Zhang, Gaimei AU - Li, Ming AU - Wu, Jing AU - Zhu, Hongfei 
JO  - Journal of Animal and Veterinary Advances
VL  - 13
IS  - 8
SP  - 549
EP  - 557
PY  - 2014
DA  - 2001/08/19
SN  - 1680-5593
DO  - javaa.2014.549.557
UR  - https://makhillpublications.co/view-article.php?doi=javaa.2014.549.557
KW  - Bovine tuberculosis
KW  -TB10.4
KW  -T cell activity
KW  -B cell activity
KW  -the gamma interferon (IFN-Y) release assay
AB  - Bovine tuberculosis caused mainly by <I>Mycobacterium bovis</I> 
  is a serious zoonosis in the world that can infect cattle and humans as well. 
  Prevention and eradication of this disease require effective and sensitive diagnosis 
  method. While the traditional tuberculin skin test and IFN-&#947; release assay 
  stimuli as PPD-B may share common antigens with <I>Mycobacterium avium</I> which 
  could cause false positive results. To overcome this problem, studies are focus 
  on screening more specific proteins to replace PPD for stimuli recently. Previous 
  research showed that TB10.4 could activate T cell stimulation therefore, researchers 
  optimized the expression of TB10.4 by comparing three different recombinant 
  proteins and using as supplementary antigen stimuli was detected by in diagnosis 
  of bovine tuberculosis. The pET-32a (+) plasmid and pET-28a (+) plasmid were 
  chosen which are different in the solubility of the recombinant protein. After 
  transformation, expression and purification of the recombinants, the recombinant 
  proteins rTB10.4-1 and rTB10.4-3 were obtained successfully. In order to analysis 
  the importance of the C terminal for the activity of TB10.4, the initiation 
  and termination codon were also introduced into the N and C terminal of <I>TB10.4</I> 
  gene and the rTB10.4-2 lack of His tag protein in the C terminal was obtained. 
  The result of SDS-PAGE showed that rTB10.4-1, TB10.4-2 and rTB10.4-3 were 30, 
  30 and 12 ku, respectively which were just the same size as expected. The biological 
  activities of these recombinant-proteins were compared by IFN-&#947; release 
  assay, real-time PCR and Western blot and the results showed that all of the 
  proteins had good T cell activity and B cell activity. In comparison, rTB10.4-1 
  possessed strongest activity while rTB10.4-3 was the lowest one as the inclusion 
  body. The skin test on guinea pigs showed that the DTH in <I>M. bovis</I>-infected 
  group could be detected intensively by using mixture of recombinant proteins 
  rTB10.4-1 and rCE in skin test compared to control group. In summary, the recombinant 
  protein of TB10.4 could be obtained successfully and be used as antigen stimuli 
  in the skin test which would lay the foundation for manufacture of the diagnosis 
  kit for bovine tuberculosis with characteristics of high specificity, sensitivity 
  and reproducibility.
ER  -