Question

what does the enzyme phosphatase do in regards to nucleic acid biochemistry

Answer 1

Role of phosphatase (alkaline and Antarctic phosphate ) in biochemistry of nucleic is discussed below:

1. Antarctic Phosphatase catalyzes the removal of 5´ phosphate from DNA and RNA. Since phosphatase-treated fragments lack the 5´ phosphoryl termini required by ligases, they cannot self-ligate (1). hence, is used to decrease the vector background in various cloning strategies.

Roles in nucleic acid:

• Removing 5´ phosphates from DNA, and RNA
• Prevention of recircularization of cloning vectors
• Dephosphorylation of proteins
• Preparation of templates for 5´ end labeling
• Removal of dNTPs and pyrophosphate from PCR reactions

1. Alkaline Phosphatase, Calf Intestinal (CIP) nonspecifically catalyzes the dephosphorylation of 5´ and 3´ ends of DNA and RNA phosphomonoesters. It also, hydrolyses ribo-, as well as deoxyribonucleoside triphosphates (NTPs and dNTPs). CIP is useful in many molecular biology applications such as the removal of phosphorylated ends of DNA and RNA for subsequent use in cloning or end-labeling of probes. In cloning, dephosphorylation prevents religation of linearized plasmid DNA. The enzyme acts on 5´ protruding, 5´ recessed and blunt ends. CIP may also be used to degrade unincorporated dNTPs in PCR reactions to prepare templates for DNA sequencing or SNP analysis

Various roles:

• Dephosphorylation of cloning vector DNA to prevent recircularization during ligation
• Dephosphorylation of DNA prior to end-labeling using T4 Polynucleotide Kinase
• Treatment of dNTPs in PCR reactions prior to sequencing or SNP analysis
• Dephosphorylation of DNA and RNA