A System for Rapid DNA Sequencing with Fluorescent Chain-Terminating Dideoxynucleotides

Prober, James M.; Trainor, George L.; Dam, Rudy J.; Hobbs, Frank W.; Robertson, Charles W.; Zagursky, Robert J.; Cocuzza, Anthony J.; Jensen, Mark A.; Baumeister, Kirk

doi:10.1126/science.2443975

Cited by 779 publications

(396 citation statements)

References 18 publications

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“…The fragments containing vectors were sequenced using the chain termination dideoxy method. 27 Southern blot. 9 ml of each PCR reaction were separated in a 2% agarose gel and afterwards transfered onto a nylon membrane by capillary blot.…”

Section: Glycerol-3-phosphate Dehydrogenase (Gpdh) Measurementmentioning

confidence: 99%

Evidence for a local renin angiotensin system in primary cultured human preadipocytes

et al. 1999

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OBJECTIVE: To investigate whether human preadipocytes possess a complete functional renin angiotensin system. MEASUREMENTS: Gene expression of angiotensinogen, renin, renin binding protein, angiotensin converting enzyme (ACE) and angiotensin II (ang II) receptor type 1 in human preadipocytes; ACE protein and ang II production of human adipose tissue stromal cells differentiated or not in primary culture. RESULTS: All genes mentioned above were found to be expressed in human preadipocytes. ACE was translated into protein as detected by western blot. Ang II was secreted both by undifferentiated preadipocytes and immature adipocytes, and its production was signi®cantly elevated in differentiated cells. CONCLUSIONS: Preadipocytes from human adipose tissue express a functional renin angiotensin system (RAS).

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Section: Glycerol-3-phosphate Dehydrogenase (Gpdh) Measurementmentioning

confidence: 99%

Evidence for a local renin angiotensin system in primary cultured human preadipocytes

et al. 1999

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“…Positive clones were identified from each isolate that contained inserts from 2.5 to 3.1 kb in size. The 3' 2400 bases of a 3.1 kb cDNA clone from the NADC isolate and a 3.0 kb cDNA clone of the KCD isolate were sequenced (Tabor & Richardson, 1987;Sanger et al, 1977) using an automated nucleic acid sequencer (Smith et al, 1986;Prober et al, 1987). When compared to the published sequence of the CFI/68 isolate of FCV (Neill et al, 1991), the nucleotide sequences of the NADC and KCD isolates were 81% and 79 % similar to the CFI/68 isolate.…”

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confidence: 99%

Analysis of feline calicivirus capsid protein genes: identification of variable antigenic determinant regions of the protein

Seal¹,

Ridpath

Mengeling³

1993

Journal of General Virology

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“…Furthermore, steadystate kinetic studies have identified hyperthermophilic DNA polymerase residues important for polymerization and exonuclease activities and for nucleotide binding (18,29,(31)(32)(33)(34)(35). Nucleotide analogs have also been important in identifying dNTP recognition determinants important in the polymerase reaction (32)(33)(34)(35)(36) and have proven useful in a variety of molecular biology applications, such as DNA sequencing and detection of single nucleotide polymorphisms (37)(38)(39)(40)(41). One group of analogs, 9-[(2-hydroxyethoxy)methyl]X triphosphates (where X is adenine, cytosine, guanine, or thymine; acyNTPs), 1 is particularly intriguing due to the wide spectrum of incorporation efficiency noted in different DNA polymerases, even within the same family of polymerase.…”

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Comparative Kinetics of Nucleotide Analog Incorporation by Vent DNA Polymerase

Gardner

Joyce

Jack

2004

Journal of Biological Chemistry

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Comparative kinetic and structural analyses of a variety of polymerases have revealed both common and divergent elements of nucleotide discrimination. Although the parameters for dNTP incorporation by the hyperthermophilic archaeal Family B Vent DNA polymerase are similar to those previously derived for Family A and B DNA polymerases, parameters for analog incorporation reveal alternative strategies for discrimination by this enzyme. Discrimination against ribonucleotides was characterized by a decrease in the affinity of NTP binding and a lower rate of phosphoryl transfer, whereas discrimination against ddNTPs was almost exclusively due to a slower rate of phosphodiester bond formation. Unlike Family A DNA polymerases, incorporation of 9-[(2-hydroxyethoxy)methyl]X triphosphates (where X is adenine, cytosine, guanine, or thymine; acyNTPs) by Vent DNA polymerase was enhanced over ddNTPs via a 50-fold increase in phosphoryl transfer rate. Furthermore, a mutant with increased propensity for nucleotide analog incorporation (Vent A488L DNA polymerase) had unaltered dNTP incorporation while displaying enhanced nucleotide analog binding affinity and rates of phosphoryl transfer. Based on kinetic data and available structural information from other DNA polymerases, we propose active site models for dNTP, ddNTP, and acyNTP selection by hyperthermophilic archaeal DNA polymerases to rationalize structural and functional differences between polymerases.All free living organisms encode several DNA polymerases that are jointly responsible for the replication and maintenance of their genomes, thereby ensuring accurate transmission of genetic information (1-3). The majority of identified DNA polymerases can be classified into Families A, B, C, and Y according to amino acid sequence similarities to Escherichia coli polymerases I, II, III, and IV/V, respectively (4, 5). Additional families have been identified, including the two-subunit replicative DNA polymerases from hyperthermophilic Archaea (Family D) (6) and eukaryotic DNA polymerase ␤ and terminal transferases (Family X) (4).Structural and kinetic analyses of Family A (7-14) and Family B (15-25) DNA polymerases have increased the understanding of nucleotide selection and incorporation mechanisms. Although amino acid sequences diverge between these two families, the structures of Family A and B DNA polymerases share recognizable finger, thumb, and palm subdomains that allow comparison of structural elements important for function (3, 11). In the case of Family A DNA polymerases from bacteriophage T7, Escherichia coli (Klenow fragment, large fragment of DNA polymerase I), and Thermus aquaticus, as well as the Family B DNA polymerase from bacteriophage RB69, interpretation of the structural information is complemented by steady-state and pre-steady-state kinetic studies, allowing a detailed description of the polymerization pathway. Reaction parameters describing the discrimination against naturally occurring nucleotide analogs encountered in vivo, such as NTPs, or unnatural n...

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A System for Rapid DNA Sequencing with Fluorescent Chain-Terminating Dideoxynucleotides

Cited by 779 publications

References 18 publications

Evidence for a local renin angiotensin system in primary cultured human preadipocytes

Evidence for a local renin angiotensin system in primary cultured human preadipocytes

Analysis of feline calicivirus capsid protein genes: identification of variable antigenic determinant regions of the protein

Comparative Kinetics of Nucleotide Analog Incorporation by Vent DNA Polymerase

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