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Short Communication · Volume 4, Issue 7, Pages 363–369, 8 July 2016 · Open Access

Genetic Characterization and Expression Analysis of Recombinant Manganese Superoxide Dismutase (MnSOD) from Spontaneously Occurring Canine Mammary Tumor

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Journal:
Advances in Animal and Veterinary Sciences
Published:
8 July 2016
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From the article

  • Keywords | MnSOD, Sequence characterization, Dog mammary cancer, Expression.
  • The recombinant protein may prove useful in determining effect of MnSOD on dog cancer cells and also in development of assays for detecting MnSOD auto-antibodies in cases of dog cancers.
  • Comparison of the deduced amino acid sequences from dog protein with human MnSOD revealed 92% similarity between the two (Figure 3).
  • Upon immunoblot analysis, band corresponding to the size of recombinant dog MnSOD was observed which shows immunoreactivity of the dog protein with commercial polyclonal antibody against human SOD2 protein.

Selected from the article's own text.

Abstract

Richa Arora1, Sonal Saxena1*, Sameer Shrivastava1, Shahid Hussain1, Saumya Sirivastava1, Resma Kochumanayin Vasu1, Priyanka Sharma1, monalisa Sahoo2, Naveen Kumar3

1Division of Veterinary Biotechnology, ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, UP, India; 2Division of Veterinary Pathology, ICAR-Indian Veterinary Research Institute, Izatnagar Bareilly, UP, India; 3Di- vision of Veterinary Surgery, ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, UP, India.

Abstract | Cancer onset typically encompasses alterations in cellular proliferation, production of reactive oxygen spe- cies (ROS), and DNA damage. Manganese superoxide dismutase (MnSOD) is one of the key enzymes for controlling oxidative stress. e role of MnSOD in cancer development is complex. Although initially considered as a tumour sup- pressor protein, recently it has been looked upon as a potential marker for cancer metastasis. ough MnSOD has been studied extensively in human cancers, not much work has been done for elucidating its role in dog cancers. Further dog MnSOD gene has not been sequenced so far, although dog MnSOD gene sequence predicted using computational biology approaches is present in the GenBank database. erefore in this study, full-length ORF of sod2 gene, a gene encoding MnSOD enzyme, from a case of canine mixed mammary capillary cystic adenocarcinoma was cloned and sequenced. e sequence showed 100% similarity with the predicted dog MnSOD sequence present in GenBank data- base based on computational algorithms. Comparison of deduced amino acid sequence from Canis lupus familiaris and Homo sapiens revealed 92% similarity, con rming the conserved nature of the protein. Further the full-length MnSOD was expressed in E.coli and puri ed protein was isolated successfully. e recombinant MnSOD from dog o ers a new tool for studying e ect of the anti-oxidant enzyme on dog cancer cells, wound healing and other disorders.

Acknowledgements

The authors are thankful to Director, IVRI, and Izatnagar for providing necessary infrastructure and facilities to carry out this research work. Authors are also thankful to Department of Biotechnology (DBT), Govt. of India (Project grant No. BT/PR13900/ADV/57/44/2010) and Indian Council of Agricultural Research (ICAR) NAE programme on biosensors for supporting the study by research grants.

Conflict of Interest

Authors of this manuscript declare no conflict of interest.

Authors’ Contribution

R. Arora conducted the experiments. S. Saxena & S. Shrivastava designed the work, helped in conducting the experiments and drafted the manuscript. S. Hussain, R.K. Vasu helped in optimization of MnSOD protein expression. S. Srivastava & P. Sharma helped in data analysis. M.Sahoo helped in histopathological analysis. N. Kumar collected tumour tissues from surgically operated CMT cases.

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Genetic Characterization and Expression Analysis of Recombinant Manganese Superoxide Dismutase (MnSOD) from Spontaneously Occurring Canine Mammary Tumor · ResearchersLinks