Chimeric protein alpha BNAC crystallin with extraordinarily high chaperone-like activity and a method thereof
AssigneeCouncil of Scientific and Industrial Research
InventorsL.V. Siva Kumar, Chandra Mohan Rao
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Abstract
The present invention relates to a chimera α BNAC of SEQ ID No.1 and a corresponding chimeric protein α BNAC crystallin of SEQ ID No. 2, with said chimeric protein having extraordinarily high chaperone-like activity and a method of preventing protein aggregation using said chimeric protein.
Claims32
A chimera α BNAC of SEQ ID No. 1.
A method of producing chimera α BNAC crystallin of SEQ ID No. 1 and a chimeric protein α BNAC crystallin thereof having extraordinarily high chaperone-like activity, said method comprising:
- (a) excising exon 1 of α A crystallin coding for N-terminal domain of eye lens protein α A crystallin using restriction enzyme XmnI,
- (b) excising exon 1 of α B crystallin coding for N-terminal domain of eye lens protein α B crystallin using restriction enzyme XmnI,
- (c) substituting said excised exon 1 of α A crystallin with excised exon 1 of α B crystallin in α A crystallin,
- (d) obtaining chimera α BNAC crystallin,
- (e) expressing said α BNAC crystallin in E. Coli, and
- (f) purifying expressed chimeric protein α BNAC crystallin.
A method of using chimeric protein α BNAC crystallin of SEQ ID 2 for preventing protein aggregation, said method comprising:
- (a) mixing test protein with said chimeric protein,
- (b) incubating mixture at room temperature,
- (c) adding protein denaturing agent to the mixture, and
- (d) estimating protein aggregation (if any) using conventional methods.
Dependencies are inferred from the claim text.