Janak Dhakal - Academia.edu (original) (raw)
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CSIC (Consejo Superior de Investigaciones Científicas-Spanish National Research Council)
Hajee Mohammad Danesh Science & Technolgy University, Bangladesh
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Papers by Janak Dhakal
The Indian Veterinary Journal, Oct 1, 2013
The Indian veterinary journal
Journal of Crop Improvement
Indian journal of medical microbiology
Heat shock proteins (Hsps) are evolutionary ancient and highly conserved molecular chaperons foun... more Heat shock proteins (Hsps) are evolutionary ancient and highly conserved molecular chaperons found in prokaryotes as well as eukaryotes. Hsp70 is a predominant member of Hsp family. Microbial Hsp70s (mHsp70s) have acquired special significance in immunity since they have been shown to be potent activators of the innate immune system and generate specific immune responses against tumours and infectious agents. The present study was aimed to clone express and purify recombinant Hsp70 from the Mycobacterium tuberculosis and characterise it immunologically. The study also aimed at determining the potential of recombinant M. tuberculosis heat shock protein (rMTB-Hsp70) as adjuvant or antigen carrier. Cloning of M. tuberculosis heat shock protein (MTB-Hsp70) amplicon was carried out using the pGEMT-Easy vector although for expression, pProExHTb prokaryotic expression vector was used. Purification of recombinant Hsp70 was carried out by nickel-nitrilotriacetic acid (Ni-NTA) affinity chroma...
Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 2006
Growth hormone is an essential polypeptide required for normal growth and development of vertebra... more Growth hormone is an essential polypeptide required for normal growth and development of vertebrates. The pejerrey fish, Odontesthes bonariensis, is a South American atherinid freshwater fish considered as a promising species for aquaculture. Although growth hormone has been characterized in a number of fish, there are no published data on the structure of this hormone in atherinids, except that of a related species Odontesthes argentinensis. In this paper, the molecular cloning, expression and immunological characterization of pejerrey growth hormone (pjGH) is described. The predicted amino acid sequence of pjGH cDNA consisted of 204 amino acid residues with an estimated molecular mass of 23 kDa. Amino acid sequence was highly conserved among the two Atheriniformes where the growth hormone sequences are known (99% aa identity), highly to moderately conserve (75-92% aa identity) when compared to the other members of Acantopterigii superorder and clearly less conserved (49-66% identity) when compared to Salmoniformes (Protacanthopterygii), Cypriniformes and Siluriformes (Ostariophysi). A phylogenetic tree depicting the relationship of various teleost GH nucleotide sequences was inferred. Pejerrey GH was produced using recombinant DNA technology in a bacterial system, representing the first time an atherinid growth hormone protein was expressed as a recombinant protein in Escherichia coli. A specific antiserum of this hormone was raised in rabbits and its specificity tested by using Western blot and immunocytochemistry. The distribution of pjGH mRNA was also studied by RT-PCR and Southern blot analysis. The transcript was detected not only in the pituitary gland but also in the testis.
The Indian Veterinary Journal, Oct 1, 2013
The Indian veterinary journal
Journal of Crop Improvement
Indian journal of medical microbiology
Heat shock proteins (Hsps) are evolutionary ancient and highly conserved molecular chaperons foun... more Heat shock proteins (Hsps) are evolutionary ancient and highly conserved molecular chaperons found in prokaryotes as well as eukaryotes. Hsp70 is a predominant member of Hsp family. Microbial Hsp70s (mHsp70s) have acquired special significance in immunity since they have been shown to be potent activators of the innate immune system and generate specific immune responses against tumours and infectious agents. The present study was aimed to clone express and purify recombinant Hsp70 from the Mycobacterium tuberculosis and characterise it immunologically. The study also aimed at determining the potential of recombinant M. tuberculosis heat shock protein (rMTB-Hsp70) as adjuvant or antigen carrier. Cloning of M. tuberculosis heat shock protein (MTB-Hsp70) amplicon was carried out using the pGEMT-Easy vector although for expression, pProExHTb prokaryotic expression vector was used. Purification of recombinant Hsp70 was carried out by nickel-nitrilotriacetic acid (Ni-NTA) affinity chroma...
Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 2006
Growth hormone is an essential polypeptide required for normal growth and development of vertebra... more Growth hormone is an essential polypeptide required for normal growth and development of vertebrates. The pejerrey fish, Odontesthes bonariensis, is a South American atherinid freshwater fish considered as a promising species for aquaculture. Although growth hormone has been characterized in a number of fish, there are no published data on the structure of this hormone in atherinids, except that of a related species Odontesthes argentinensis. In this paper, the molecular cloning, expression and immunological characterization of pejerrey growth hormone (pjGH) is described. The predicted amino acid sequence of pjGH cDNA consisted of 204 amino acid residues with an estimated molecular mass of 23 kDa. Amino acid sequence was highly conserved among the two Atheriniformes where the growth hormone sequences are known (99% aa identity), highly to moderately conserve (75-92% aa identity) when compared to the other members of Acantopterigii superorder and clearly less conserved (49-66% identity) when compared to Salmoniformes (Protacanthopterygii), Cypriniformes and Siluriformes (Ostariophysi). A phylogenetic tree depicting the relationship of various teleost GH nucleotide sequences was inferred. Pejerrey GH was produced using recombinant DNA technology in a bacterial system, representing the first time an atherinid growth hormone protein was expressed as a recombinant protein in Escherichia coli. A specific antiserum of this hormone was raised in rabbits and its specificity tested by using Western blot and immunocytochemistry. The distribution of pjGH mRNA was also studied by RT-PCR and Southern blot analysis. The transcript was detected not only in the pituitary gland but also in the testis.