124
4.11 Conclusions
Enzymes have a pivotal role in molecular biology and a wide array of enzymes is
now commercially available for cleaving, ligating, synthesis, and modification of
nucleic acids. A majority of these enzymes are produced as recombinant proteins
and expressed in heterologous hosts by DNA cloning. Enzymes can be even modified to suit specific needs by altering coding sequences to enhance its activity or to
remove a particular catalytic activity. Thus, a good understanding of the various
enzymes available along with their catalytic characteristics and optimum conditions
is essential to select the best one for a particular requirement.
References
Alsmadi O, Alkayal F, Monies D, Meyer BF (2009) Specific and complete human genome amplification with improved yield achieved by phi29 DNA polymerase and a novel primer at elevated
temperature. BMC Res Note 2:48
Anfinsen CB, Haber E, Sela M, White FH (1961) The kinetics of formation of native ribonuclease
during oxidation of the reduced polypeptide chain. Proc Natl Acad Sci U S A 47(9):1309–1314
Angers M, Cloutier JF, Castonguay A, Drouin R (2001) Optimal conditions to use Pfu exo(-) DNA
polymerase for highly efficient ligation-mediated polymerase chain reaction protocols. Nucleic
Acids Res 29:E83
Baluda MA, Perbal B, Rushlow KE, Papas TS (1983) Avian myeloblastosis virus: a model for the
generation of viral oncogenes from potentially oncogenic cellular genetic elements. Folia Biol
29:18–34
Belkin S, Jannasch HW (1985) A new extremely thermophilic, sulfur-reducing heterotrophic,
marine bacterium. Arch Microbiol 141(3):181–186
Benedik MJ, Strych U (1998) Serratia marcescens and its extracellular nuclease. FEMS Microbiol
Lett 165:1–13
Blanco L, Bernad A, Lazaro JM, Martin G, Garmendia C, Salas M (1989) Highly efficient DNA
synthesis by the phage phi 29 DNA polymerase. Symmetrical mode of DNA replication. J Biol
Chem 264:8935–8940
Blondal T, Hjorleifsdottir SH, Fridjonsson OF, Ævarsson A, Skirnisdottir S, Hermannsdottir AG,
Kristjansson JK (2003) Discovery and characterization of a thermostable bacteriophage RNA
ligase homologous to T4 RNA ligase 1. Nucleic Acids Res 31(24):7247–7254
Blondal T, Thorisdottir A, Unnsteinsdottir U, Hjorleifsdottir S, Ævarsson A, Ernstsson S,
Kristjansson JK (2005) Isolation and characterization of a thermostable RNA ligase 1 from a
Thermus scotoductus bacteriophage TS2126 with good single-stranded DNA ligation properties. Nucleic Acids Res 33(1):135–142
Brock TD, Freeze H (1969) Thermus aquaticus gen. n. and sp. n., a nonsporulating extreme thermophile. J Bacteriol 98:289–297
Burgers PM, Koonin EV, Bruford E, Blanco L, Burtis KC, Christman MF, Copeland WC, Friedberg
EC, Hanaoka F, Hinkle DC, Lawrence CW, Nakanishi M, Ohmori H, Prakash L, Prakash S,
Reynaud CA, Sugino A, Todo T, Wang Z, Weill JC, Woodgate R (2001) Eukaryotic DNA polymerases: proposal for a revised nomenclature. J Biol Chem 276:43487–43490
Butler ET, Chamberlin MJ (1982) Bacteriophage SP6-specific RNA polymerase I. Isolation and
characterization of the enzyme. J Biol Chem 257(10):5772–5778
Cai L, Hu C, Shen S, Wang W, Huang W (2004) Characterization of bacteriophage T3 DNA ligase.
J Biochem 135(3):397–403
G. Valsala and S. Sugathan
4.11 Conclusions
Enzymes have a pivotal role in molecular biology and a wide array of enzymes is
now commercially available for cleaving, ligating, synthesis, and modification of
nucleic acids. A majority of these enzymes are produced as recombinant proteins
and expressed in heterologous hosts by DNA cloning. Enzymes can be even modified to suit specific needs by altering coding sequences to enhance its activity or to
remove a particular catalytic activity. Thus, a good understanding of the various
enzymes available along with their catalytic characteristics and optimum conditions
is essential to select the best one for a particular requirement.
References
Alsmadi O, Alkayal F, Monies D, Meyer BF (2009) Specific and complete human genome amplification with improved yield achieved by phi29 DNA polymerase and a novel primer at elevated
temperature. BMC Res Note 2:48
Anfinsen CB, Haber E, Sela M, White FH (1961) The kinetics of formation of native ribonuclease
during oxidation of the reduced polypeptide chain. Proc Natl Acad Sci U S A 47(9):1309–1314
Angers M, Cloutier JF, Castonguay A, Drouin R (2001) Optimal conditions to use Pfu exo(-) DNA
polymerase for highly efficient ligation-mediated polymerase chain reaction protocols. Nucleic
Acids Res 29:E83
Baluda MA, Perbal B, Rushlow KE, Papas TS (1983) Avian myeloblastosis virus: a model for the
generation of viral oncogenes from potentially oncogenic cellular genetic elements. Folia Biol
29:18–34
Belkin S, Jannasch HW (1985) A new extremely thermophilic, sulfur-reducing heterotrophic,
marine bacterium. Arch Microbiol 141(3):181–186
Benedik MJ, Strych U (1998) Serratia marcescens and its extracellular nuclease. FEMS Microbiol
Lett 165:1–13
Blanco L, Bernad A, Lazaro JM, Martin G, Garmendia C, Salas M (1989) Highly efficient DNA
synthesis by the phage phi 29 DNA polymerase. Symmetrical mode of DNA replication. J Biol
Chem 264:8935–8940
Blondal T, Hjorleifsdottir SH, Fridjonsson OF, Ævarsson A, Skirnisdottir S, Hermannsdottir AG,
Kristjansson JK (2003) Discovery and characterization of a thermostable bacteriophage RNA
ligase homologous to T4 RNA ligase 1. Nucleic Acids Res 31(24):7247–7254
Blondal T, Thorisdottir A, Unnsteinsdottir U, Hjorleifsdottir S, Ævarsson A, Ernstsson S,
Kristjansson JK (2005) Isolation and characterization of a thermostable RNA ligase 1 from a
Thermus scotoductus bacteriophage TS2126 with good single-stranded DNA ligation properties. Nucleic Acids Res 33(1):135–142
Brock TD, Freeze H (1969) Thermus aquaticus gen. n. and sp. n., a nonsporulating extreme thermophile. J Bacteriol 98:289–297
Burgers PM, Koonin EV, Bruford E, Blanco L, Burtis KC, Christman MF, Copeland WC, Friedberg
EC, Hanaoka F, Hinkle DC, Lawrence CW, Nakanishi M, Ohmori H, Prakash L, Prakash S,
Reynaud CA, Sugino A, Todo T, Wang Z, Weill JC, Woodgate R (2001) Eukaryotic DNA polymerases: proposal for a revised nomenclature. J Biol Chem 276:43487–43490
Butler ET, Chamberlin MJ (1982) Bacteriophage SP6-specific RNA polymerase I. Isolation and
characterization of the enzyme. J Biol Chem 257(10):5772–5778
Cai L, Hu C, Shen S, Wang W, Huang W (2004) Characterization of bacteriophage T3 DNA ligase.
J Biochem 135(3):397–403
G. Valsala and S. Sugathan