| 摘要 |
The present invention relates to polypeptides, compositions and methods for enhancing synthesis of nucleic acid molecules. In a preferred aspect, the invention relates to inhibition or control of nucleic acid synthesis, sequencing or amplification. Specifically, the present invention discloses polypeptides having affinity for double-stranded and/or single-stranded nucleic acid molecules and/or single-stranded/double-stranded nucleic acid complexes (e.g., primer/template complexes, double-stranded templates, single-stranded templates or single-stranded primers) for use in such enhanced synthesis and more particularly to polymerases having reduced polymerase and optionally reduced exonuclease activities (3' to 5' and/or 5' to 3' exonuclease activity), and to nucleases having reduced nuclease activity. The polypeptides of the invention are capable of inhibiting nonspecific nucleic acid synthesis at ambient temperature. Thus, in a preferred aspect, the invention relates to "hot start" synthesis of nucleic acid molecules. Accordingly, the invention prevents non-specific nucleic acid synthesis at low temperatures, for example during reaction set up. The invention also relates to kits for synthesizing, amplifying, reverse transcribing or sequencing nucleic acid molecules comprising one or more of the polypeptides or compositions of the invention. The invention also relates to compositions prepared for carrying out the methods of the invention and to compositions made after or during such methods. The invention also generally relates to polypeptides and compositions useful for inhibiting or preventing degradation of various nucleic acid molecules.
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