Enhanced protein thermostability and temperature resistance
AssigneeUniversity of Maryland Biotechnology Institute
InventorsFrank T. Robb, Pongpan Laksanalamai
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Abstract
Small heat shock proteins, e.g., Pyrococcus fuiosus (Pfu-sHSP and/or Pfu-tsHSP), confer thermotolerance on cellular cultures and on proteins in cellular extracts during prolonged incubation at elevated temperature, demonstrating the ability to protect cellular proteins and maintain cellular viability under heat stress conditions. Such heat shock proteins are effective to combat enzymatic aggregation and intracellular precipitation during heat stress, and thereby enable enhancement of the utility and stability of enzymes in various applications, e.g., Taq polymerase in PCR applications, digestive enzymes in microbial degradative applications, etc.
Claims33
A purified and isolated nucleic acid sequence encoding a heat shock protein, consisting of SEQ ID NO:
- 1.
An isolated protein encoded by the nucleic acid consisting of SEQ ID NO. 5.
An isolated protein consisting of the amino acid sequence of SEQ ID NO. 6.
A composition comprising (i) a biological component and (ii) a heat shock protein (HSP) consisting of the amino acid sequence of SEQ ID NO. 6.
A PCR kit including PCR primers, Taq polymerase, deoxyribonucleoside triphoephates and a heat shock protein (HSP) consisting of the amino acid sequence of SEQ ID NO. 6.
A method of stabilizing a protein solution, including a first protein therein, against hear-mediated agglomeration of said first protein in the solution, comprising incorporating in the solution a heat shock protein that is non-endogenous with respect to said first protein, wherein said heat shock protein consisting of the amino acid sequence of SEQ ID N0.6.
A PCR kit, comprising a sHSP19 heat shock protein, and optionally including at least one component selected from the group consisting of (i) PCR primers, (ii) a polymerase effective for PCR and (iii) deoxyribonucleoside triphosphates, wherein the heat shock protein consists of SEQ ID NO:
- 6.
A recombinant DNA vector for expression of Pfu-tsHSP, wherein the recombinant DNA vector comprises a nucleic acid consisting of SEQ ID NO. 5.
A transformed cell capable of expressing Pfu-tsHSP, wherein the transformed cell comprises a recombinant DNA vector comprising a nucleic acid consisting of SEQ ID NO. 5.
A method of stabilizing a medium including a biological component susceptible to heat-mediated degradation, comprising incorporating in said medium an effective amount of HSP consisting of the amino mold sequence of SEQ ID NO:
- 6.
An isolated protein comprising the amino acid sequence of SEQ ID NO. 6, wherein the end five amino acid residues at the carboxyl terminal consist of Lys His Pro Thr Lys.
Dependencies are inferred from the claim text.