Essential for the replication of viral ssDNA. The closed circular ssDNA genome is first converted to a superhelical dsDNA. Rep binds a specific region at the genome origin of replication. It introduces an endonucleolytic nick within the conserved sequence 5-TAATATTAC-3 in the intergenic region of the genome present in all giniviruses, thereby initiating the rolling circle replication (RCR). Following cleavage, binds covalently to the 5-phosphate of DNA as a tyrosyl ester. The cleavage gives rise to a free 3-OH that serves as a primer for the cellular DNA polymerase. The polymerase synthesizes the (+) strand DNA by rolling circle mechanism. After one round of replication, a Rep-catalyzed nucleotidyl transfer reaction releases a circular single-stranded virus genome, thereby terminating the replication. Displays origin-specific DNA cleavage, nucleotidyl transferase, ATPase and helicase activities. Source: Recombinant protein corresponding to aa1-358 from african cassava mosaic virus Replication-associated Protein, fused to His-SUMO-Tag at N-terminal, expressed in E. coli. Molecular Weight: ~56.3kD Amino Acid Sequence: MRTPRFRIQAKNVFLTYPKCSIPKEHLLSFIQTLSLQSNPKFIKICRELHQNGEPHLHALIQFEGKITITNNRLFDCVHPSCSTSFHPNIQGAKSSSDVKSYLDKDGDTVEWGQFQIDGRSARGGQQSANDAYAKALNSGSKSEALNVIRELVPKDFVLQFHNLNSNLDRIFQEPPAPYVSPFPCSSFDQVPVEIEEWVADNVRDSAARPWRPNSIVIEGDSRTGKTIWARSLGPHNYLCGHLDLSPKVFNNAAWYNVIDDVDPHYLKHFKEFMGSQRDWQSNTKYGKPVQIKGGIPTIFLCNPGPTSSYKEFLAEEKQEALKAWALKNAIFITLTEPLYSGSNQSHSQTSQEASHPA Storage and Stability: Lyophilized and reconstituted products are stable for 6 months after receipt at -20C. Reconstitute with sterile ddH2O. Aliquot to avoid repeated freezing and thawing. Store at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Supplied as a lyophilized powder from 20mM Tris-HCl, 0.5M sodium chloride, pH 8.0, 6% trehalose. Reconstitute with sterile ddH2O to a concentration of 0.1-1mg/ml.
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