A method for filling in the cohesive ends of double-stranded DNA using Pfu DNA polymerase

2005 ◽  
Vol 42 (3) ◽  
pp. 223 ◽  
Author(s):  
Xin Li ◽  
Shaohui Yang ◽  
Dongfeng Ding ◽  
Jianhua Hou ◽  
Zhaoxia Jin ◽  
...  
Keyword(s):  
Dna Polymerase ◽  
Double Stranded Dna ◽  
Pfu Dna Polymerase

Oligo(A)-stimulated Tetrahymena rDNA synthesis in vitro

1981 ◽  
Vol 59 (6) ◽  
pp. 396-403 ◽  
Author(s):  
Peter R. Ganz ◽  
Gyorgy B. Kiss ◽  
Ronald E. Pearlman
Keyword(s):  
Rna Polymerase ◽  
Dna Synthesis ◽  
Dna Polymerase ◽  
Ribosomal Rna ◽  
Replication Proteins ◽  
General Applicability ◽  
In Vitro Synthesis ◽  
Restriction Fragments ◽  
Double Stranded Dna

The synthesis of Tetrahymena rDNA has been examined using purified DNA polymerase and partially purified preparations of homologous replication enzymes (fraction IV). DNA synthesis with purified DNA polymerase alone was less than that with fraction IV enzymes. This suggested that there were additional factors in fraction IV other than DNA polymerase which contributed to or enhanced rDNA synthesis in vitro. Neither hybridization of rDNA with Tetrahymena ribosomal RNA nor preincubation of rDNA with homologous or heterologous RNA polymerase served to stimulate in vitro synthesis by fraction IV enzymes. However, when rDNA was hybridized with oligoriboadenylate, DNA synthesis using fraction IV was stimulated approximately 4- to 4.5-fold over 150 min of incubation, relative to a similarly treated but unhybridized rDNA control. Using oligoriboadenylate-hybridized EcoR1 and HindIII restriction fragments of rDNA to localize the synthesis most of the in vitro synthesis occurred within a 2.4 × 106 Mr fragment encompassing the centre of the rDNA molecule. The approach of hybridizing a synthetic homooligoribonucleotide primer to double-stranded DNA should prove to be of general applicability in designing similar template–primers in other systems for the purpose of isolating replication proteins.

scholarly journals Algal Viruses with Distinct Intraspecies Host Specificities Include Identical Intein Elements

2005 ◽  
Vol 71 (7) ◽  
pp. 3599-3607 ◽  
Author(s):  
Keizo Nagasaki ◽  
Yoko Shirai ◽  
Yuji Tomaru ◽  
Kensho Nishida ◽  
Shmuel Pietrokovski
Keyword(s):  
Dna Polymerase ◽  
Homing Endonuclease ◽  
Sister Group ◽  
Dna Virus ◽  
Single Nucleotide Change ◽  
Double Stranded Dna ◽  
Endonuclease Domain ◽  
Algal Viruses ◽  
Molecular Phylogenetic ◽  
Position Coding

ABSTRACT Heterosigma akashiwo virus (HaV) is a large double-stranded DNA virus infecting the single-cell bloom-forming raphidophyte (golden brown alga) H. akashiwo. A molecular phylogenetic sequence analysis of HaV DNA polymerase showed that it forms a sister group with Phycodnaviridae algal viruses. All 10 examined HaV strains, which had distinct intraspecies host specificities, included an intein (protein intron) in their DNA polymerase genes. The 232-amino-acid inteins differed from each other by no more than a single nucleotide change. All inteins were present at the same conserved position, coding for an active-site motif, which also includes inteins in mimivirus (a very large double-stranded DNA virus of amoebae) and in several archaeal DNA polymerase genes. The HaV intein is closely related to the mimivirus intein, and both are apparently monophyletic to the archaeal inteins. These observations suggest the occurrence of horizontal transfers of inteins between viruses of different families and between archaea and viruses and reveal that viruses might be reservoirs and intermediates in horizontal transmissions of inteins. The homing endonuclease domain of the HaV intein alleles is mostly deleted. The mechanism keeping their sequences basically identical in HaV strains specific for different hosts is yet unknown. One possibility is that rapid and local changes in the HaV genome change its host specificity. This is the first report of inteins found in viruses infecting eukaryotic algae.

scholarly journals Engineered split in Pfu DNA polymerase fingers domain improves incorporation of nucleotide  -phosphate derivative

2010 ◽  
Vol 39 (5) ◽  
pp. 1801-1810 ◽  
Author(s):  
C. J. Hansen ◽  
L. Wu ◽  
J. D. Fox ◽  
B. Arezi ◽  
H. H. Hogrefe
Keyword(s):  
Dna Polymerase ◽  
Pfu Dna Polymerase

An Improved Phage Display Antibody Cloning System Using Newly Designed PCR Primers Optimized for Pfu DNA Polymerase

1995 ◽  
Vol 117 (6) ◽  
pp. 1218-1227 ◽  
Author(s):  
Hachiro I. Yamanaka ◽  
Yasuyuki Kirii ◽  
Hiroshi Ohmoto
Keyword(s):  
Phage Display ◽  
Dna Polymerase ◽  
Pcr Primers ◽  
Cloning System ◽  
Pfu Dna Polymerase ◽  
Phage Display Antibody

Double-stranded DNA binding properties of Saccharomyces cerevisiae DNA polymerase ɛ and of the Dpb3p-Dpb4p subassembly

2003 ◽  
Vol 8 (11) ◽  
pp. 873-888 ◽  
Author(s):  
Toshiaki Tsubota ◽  
Satoko Maki ◽  
Hajime Kubota ◽  
Akio Sugino ◽  
Hisaji Maki
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Binding ◽  
Dna Polymerase ◽  
Binding Properties ◽  
Double Stranded Dna ◽  
Dna Binding Properties

Purification of recombinant Pfu DNA polymerase using a new JK110 resin

2006 ◽  
Vol 23 (4) ◽  
pp. 607-609 ◽  
Author(s):  
Zhanghui Sun ◽  
Jin Cai
Keyword(s):  
Dna Polymerase ◽  
Pfu Dna Polymerase ◽  
Jk110 Resin

DNA polymerase .delta. holoenzyme: action on single-stranded DNA and on double-stranded DNA in the presence of replicative DNA helicases

Biochemistry ◽  
1995 ◽  
Vol 34 (15) ◽  
pp. 5003-5010 ◽  
Author(s):  
Vladimir N. Podust ◽  
Larissa M. Podust ◽  
Friedemann Mueller ◽  
Ulrich Huebscher
Keyword(s):  
Dna Polymerase ◽  
Dna Polymerase Delta ◽  
Single Stranded Dna ◽  
Dna Helicases ◽  
Double Stranded Dna

Molecular cloning and expression of Pfu DNA polymerase gene and its application in long-distance PCR

1998 ◽  
Vol 43 (10) ◽  
pp. 863-867 ◽  
Author(s):  
Bing Li ◽  
Cheng Gu ◽  
Jindong Zhao
Keyword(s):  
Molecular Cloning ◽  
Dna Polymerase ◽  
Cloning And Expression ◽  
Polymerase Gene ◽  
Long Distance ◽  
Dna Polymerase Gene ◽  
Long Distance Pcr ◽  
Pfu Dna Polymerase
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