Open Access
Copyright: The authors. This article is an open access
article licensed under the terms of the Creative Commons Attribution License
(http://creativecommons.org/licenses/by/2.0) which permits unrestricted use,
distribution and reproduction in any medium, provided the work is properly
cited.
Research
(Published
online: 15-12-2014)
10.
Cloning and sequence analysis of hsf,
an outer membrane protein gene of Pasteurella multocida
serotype B:2 -
A. Priyadarshini, S. Kumar, S. K. Gupta, K.
N. Viswas, R. K. Agarwal and V. P. Singh
Veterinary World, 7(12): 1090-1093
doi:
10.14202/vetworld.2014.1090-1093
A.
Priyadarshini:
Division of Veterinary Bacteriology, Indian Veterinary Research
Institute, Izatnagar, Bareilly, Uttar Pradesh, India;
adyashavet01@gmail.com
S.
Kumar:
Division of Veterinary Bacteriology, Indian Veterinary Research
Institute, Izatnagar, Bareilly, Uttar Pradesh, India;
skumar.cdri@gmail.com
S. K.
Gupta:
Division of Veterinary Bacteriology, Indian Veterinary Research
Institute, Izatnagar, Bareilly, Uttar Pradesh, India;
skgupta15@rediffmail.com
K. N.
Viswas:
Division of Veterinary Bacteriology, Indian Veterinary Research
Institute, Izatnagar, Bareilly, Uttar Pradesh, India;
vkn111@gmail.com
R. K.
Agarwal:
Division of Veterinary Bacteriology, Indian Veterinary Research
Institute, Izatnagar, Bareilly, Uttar Pradesh, India;
grace_bly@yahoo.com
V. P.
Singh: Division of Veterinary Bacteriology, Indian Veterinary
Research Institute, Izatnagar, Bareilly, Uttar Pradesh, India;
vijendra61@gmail.com
Received:
28-09-2014, Revised: 04-11-2014, Accepted: 09-11-2014 Published
online: 15-12-2014
Corresponding author:
A. Priyadarshini, email: adyashavet01@gmail.com
Abstract
Aim:
The present study was undertaken to clone, sequence and
analyze the hsf, an outer membrane protein gene of
Pasteurella multocida serotype B:2
Materials and Methods: hsf gene was amplified from
genomic DNA of P. multocida. Polymerase chain reaction (PCR)
product was cloned in pET-32a vector and was characterized. hsf
gene was sequenced, analyzed and phylogenetic tree was
constructed taking sequences of other strains.
Results: Amplicon size was found to be 785 bp. Recombinant got
characterized through colony PCR and restriction enzyme analysis.
Conclusion: hsf gene of P. multocida serotype B
is similar to serotype A, but different from serotype D. Further
work is needed to evaluate role of Hsf protein in protection
studies and to study the antigenic properties of this recombinant
protein as a candidate for vaccine.
Keywords: cloning, hsf gene, outer
membrane protein, Pasteurella multocida, phylogenetic tree,
sequencing
References
1. Boyce, J.D., Wilkie, I., Harper, M., Paustian, M.L., Kapur,
V. and Adler, B. (2002) Genomic scale analysis of Pasteurella
multocida gene expression during growth within the natural
chicken host. Infect. Immunol., 70(12): 6871-6879.
http://dx.doi.org/10.1128/IAI.70.12.6871-6879.2002
PMid:12438364 PMCid:PMC133079 |
|
|
2. Davies, R.L., MacCorquodale, R., Baillie, S. and Caffrey,
B. (2003) Characterization and comparison of Pasteurella
multocida strains associated with porcine pneumonia and
atrophic rhinitis. J. Med. Microbiol., 52: 59-67.
http://dx.doi.org/10.1099/jmm.0.05019-0
PMid:12488567 |
|
|
3. Shivachandra, S.B., Viswas, K.N. and Kumar A.A. (2011) A
review of haemorrhagic septicaemiain cattle and buffalo. Anim.
Health Res. Rev., 12(1): 67-82.
http://dx.doi.org/10.1017/S146625231100003X
PMid:21676341 |
|
|
|
4. Sarangi, L.N., Thomas, P., Gupta, S.K., Kumar, S., Viswas,
K.N. and Singh, V.P. (2014) Molecular epidemiology of
Pasteurella multocida circulating in India by multilocus
sequence typing. Transbound Emerg. Dis. |
|
|
5. Boyce, J.D., Cullen, P.A., Nguyen, V.I., Wilkie, I. and
Adler, B. (2006) Analysis of the Pasteurella multocida outer
membrane sub-proteome and its response to the in vivo
environment of the natural host. Proteomics, 6(3): 870-880.
http://dx.doi.org/10.1002/pmic.200401342
PMid:16372271 |
|
|
6. Lee, J., Kim, Y.B. and Kwon, M. (2007). Outer membrane
protein H for protective immunity against Pasteurella
multocida. J. Microbiol., 45(2): 179-184.
PMid:17483806 |
|
|
7. Hatfaludi, T.S., Al-Hasani, K., Boyce, J. D. and Adler, B.
(2010) Outer membrane proteins of Pasteurella multocida. Vet.
Microbiol., 144: 1-17.
http://dx.doi.org/10.1016/j.vetmic.2010.01.027
PMid:20197220 |
|
|
8. Hallstrom, T., Trajkovska, E., Forsgren, A. and Riesbeck,
K. (2006) Haemophilus influenzae surface fibrils contribute to
serum resistance by interacting with vitronectin. J. Immunol.,
177(1): 430-436.
http://dx.doi.org/10.4049/jimmunol.177.1.430
PMid:16785539 |
|
|
9. Paustian, M.L., May, B.J. and Kapur, V. (2002)
Transcriptional response of Pasteurella multocida to nutrient
limitation. J. Bacteriol., 184: 3734-3739.
http://dx.doi.org/10.1128/JB.184.13.3734-3739.2002
PMid:12057970 PMCid:PMC135142 |
|
|
10. Tang, X., Zhao, Z., Hu, J., Wu, B., Cai, X., He, Q. and
Chen, H. (2009) Isolation, antimicrobial resistance, and
virulence genes of Pasteurella multocida strains from swine in
China. J. Clin. Microbiol., 47(4): 951-958.
http://dx.doi.org/10.1128/JCM.02029-08
PMid:19158260 PMCid:PMC2668295 |
|
|
|
11. Sarangi, L.N., Thomas, P., Priyadarshini, A., Kumar, S.,
Viswas, K.N., Singh, V.P. and Gupta S.K. (2013) Virulence gene
profiling and antibiotic resistance pattern of Indian isolates
of Pasteurella multocida for small ruminant origin. In: XXVII
Annual Convention and National Conference of IAVMI-2013 on
Productivity Enhancement through Improved Animal Health and
Nutrition held at Lucknow, U.P. from December 13-15. p89. |
|
|
12. Shivachandra, S.B., Kumar, A., Mohanty, N.N.,
Yogisharadhya, R., Chacko, N., Viswas, K.N. and Ramakrishnan
M.A. (2014) Homogeneity of VacJ outer membrane lipoproteins
among Pasteurella multocida strains and heterogeneity among
members of Pasteurellaceae. Res. Vet. Sci., 96(3): 415-421.
http://dx.doi.org/10.1016/j.rvsc.2014.03.016
PMid:24731530 |
|
|
13. Singh, A.P., Singh, S., Ranjan, R., Gupta, S.K., Singh,
V.P. and Sharma, B. (2010) Molecular heterogeneity of plpE
gene in Indian isolates of Pasteurella multocida and
expression of recombinant PlpE in vaccine strain of
Pasteurella multocida serotype B:2. J. Vet. Sci., 11(3):
227-233.
http://dx.doi.org/10.4142/jvs.2010.11.3.227
PMid:20706030 PMCid:PMC2924484 |
|
|
14. Siju, J., Kumar, A.A., Shivachandra, S.B., Chaudhuri, P.,
Srivastava, S.K. and Singh, V.P. (2007) Cloning and
characterization of type 4 fimbrial gene (ptfA) of Pasteurella
multocida serogroup B:2 (Strain P52). Vet. Res. Commun.,
31(4): 397-404.
http://dx.doi.org/10.1007/s11259-006-3372-7
PMid:17216317 |
|
|
15. Shivachandra, S.B., Kumar, A.A., Amarnath, J., Joseph, S.,
Srivastava, S.K. and Chaudhuri, P. (2005) Cloning and
characterization of tbpA gene encoding transferrin binding
protein TbpA from Pasteurella multocida serogroup B:2 (strain
P52). Vet. Res. Commun., 29: 537-542.
http://dx.doi.org/10.1007/s11259-005-2495-6
PMid:16215844 |
|
|
|
16. Jain, A. (2013) Expression and evaluation of diagnostic
potential of recombinant outer membrane proteins of P.
multocida serotype B: 2. M.V.Sc. Thesis Submitted to Deemed
University, IVRI, Izatnagar, India. |
|
