A novel approach for high-level expression and purification of GST-fused highly thermostable Taq DNA polymerase in Escherichia coli.

Polymerases are enzymes that synthesize long chains or polymers of nucleic acids, including DNA or RNA of nucleotides. They assemble to copy the template nucleic acids DNA or RNA strand using the base-pairing interactions. One of the polymerase enzyme, Taq DNA polymerase, originally isolated from Thermus aquaticus (Taq) is an enzyme widely used in molecular biology so far.

The properties of thermostable enzymes has contributed majorly to specificity, automation, and efficacy of the polymerase chain reaction (PCR), making it a powerful tool for molecular biology research today throughout the world. Purification of Taq DNA polymerase from the original host results in low yields, labor and time consumption.

Therefore, much research has been done before to get this enzyme using an alternative host. So far, all of the existing methodology is more laborious, time consuming and requires a heavy cost. We use a new approach to purifying the enzyme with relatively high efficiency, yield and minimum time consumption using Escherichia coli (E. coli) as an alternative host.

We cloned 2500 base pairs of DNA Taq polymerase gene into pGEX-4T-1 vector containing GST-tag, downstream of tac promoter and expressed using isopropyl β-D-1-thiogalactopyranoside (IPTG) as an inducer. The enzyme was purified using chromatography efficient new approach and use in routine PCR tests in our laboratory. Our findings suggest a new approach to facilitate the availability of polymerases for molecular and diagnostic studies. In the future, may be used for purification of other recombinant peptides or proteins used in structural biology and proteomics research-based.

A novel approach for high-level expression and purification of GST-fused highly thermostable Taq DNA polymerase in Escherichia coli.
A novel approach for high-level expression and purification of GST-fused highly thermostable Taq DNA polymerase in Escherichia coli.

Outside Chelation: EDTA binds Closely Taq DNA Polymerase, Mutt and dUTPase and direct Inhibiting Activity dNTPase.

EDTA is commonly used as a metal ion chelator efficient enzyme cofactor. It is highly soluble, optically inactive and do not interfere with most chemicals used in making standard buffer EDTA common options for generating metal-free conditions for biochemical and biophysical investigations.

However, controversy in the literature about the activity of the metal-free enzyme is achieved by using EDTA or otherwise called our attention to the alleged effects of external EDTA chelation. Here, we show that EDTA compete for sites of hydrolases dUTPase nucleotide nucleotide binding by developing a network of interactions within the active site similar to the substrate.

To reach these findings, we apply the kinetics and molecular docking technique uses two different dUTPases. Furthermore, we directly measured the binding of EDTA to dUTPases and two other dNTPases, Taq polymerase and Mutt using isothermal titration calorimetry. EDTA binds proven exothermic and enthalpy driven primarily by submicromolar dissociation constant is much lower than with an enzyme: substrate or Mg: EDTA complex. Control of protein, including ATPase, do not interact with EDTA. Our findings indicate that EDTA can act as a selective inhibitor of the enzyme hydrolysis of dNTPs and urges a rethinking of the use of EDTA in enzymatic trial.

Taq DNA Polymerase

9001-500
EUR 229.2

PFU DNA Polymerase

9003-2500
EUR 614.4

PFU DNA Polymerase

9003-500
EUR 216

Laq? DNA Polymerase

9004-2500
EUR 732

Laq? DNA Polymerase

9004-500
EUR 261.6

DNA polymerase ? (Rat)

10-101 20ug
EUR 465.6
Description: The DNA polymerase ? (Rat) is available in Europe and for worldwide shipping via Gentaur.

DNA polymerase ? (Rat)

10-102 100ug
EUR 1107.6
Description: The DNA polymerase ? (Rat) is available in Europe and for worldwide shipping via Gentaur.

Taq DNA Polymerase

BA00103 500U
EUR 104.4
Description: High quality Taq polymerase for different PCR variations and downstream applications.

Taq DNA Polymerase

BA00104 1000U
EUR 147.6
Description: High quality Taq polymerase for different PCR variations and downstream applications.

Taq DNA Polymerase

BA00105 2500U
EUR 278.4
Description: High quality Taq polymerase for different PCR variations and downstream applications.

HotTaq DNA Polymerase

BA00203 500U
EUR 174
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.

HotTaq DNA Polymerase

BA00204 1000U
EUR 286.8
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.

HotTaq DNA Polymerase

BA00205 2500U
EUR 626.4
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.

RedTaq DNA Polymerase

BA00303 500U
EUR 147.6
Description: High quality RedTaq polymerase for different PCR variations and downstream applications.

RedTaq DNA Polymerase

BA00304 1000U
EUR 235.2
Description: High quality RedTaq polymerase for different PCR variations and downstream applications.

RedTaq DNA Polymerase

BA00305 2500U
EUR 495.6
Description: High quality RedTaq polymerase for different PCR variations and downstream applications.

Pfu DNA Polymerase

BA00503 500U
EUR 174
Description: High quality Pfu polymerase for different PCR variations and downstream applications.

Pfu DNA Polymerase

BA00504 1000U
EUR 286.8
Description: High quality Pfu polymerase for different PCR variations and downstream applications.

Pfu DNA Polymerase

BA00505 2500U
EUR 626.4
Description: High quality Pfu polymerase for different PCR variations and downstream applications.

BIOTAQ DNA Polymerase

BIO-21040 500 Units Ask for price

IMMOLASE DNA Polymerase

BIO-21046 250 Units Ask for price

IMMOLASE DNA Polymerase

BIO-21047 500 Units Ask for price

IMMOLASE DNA Polymerase

BIO-21048 5000 Units Ask for price

ACCUZYME DNA Polymerase

BIO-21051 250 Units Ask for price

ACCUZYME DNA Polymerase

BIO-21052 500 Units Ask for price

BIOTAQ DNA Polymerase

BIO-21060 2500 Units Ask for price

MangoTaq DNA Polymerase

BIO-21078 5000 Units Ask for price

MangoTaq DNA Polymerase

BIO-21082 2000 Units Ask for price

MangoTaq DNA Polymerase

BIO-21083 1000 Units Ask for price

VELOCITY DNA Polymerase

BIO-21098 250 Units Ask for price

VELOCITY DNA Polymerase

BIO-21099 500 Units Ask for price

MyTaq DNA Polymerase

BIO-21105 500 units Ask for price

MyTaq DNA Polymerase

BIO-21105/S Sample Ask for price

MyTaq DNA Polymerase

BIO-21106 2500 units Ask for price

MyTaq DNA Polymerase

BIO-21107 5000 units Ask for price

MyFi DNA Polymerase

BIO-21117 250 Units Ask for price

MyFi DNA Polymerase

BIO-21117/S Sample Ask for price

MyFi DNA Polymerase

BIO-21118 500 Units Ask for price

MyFi DNA Polymerase

BIO-21119 2500 Units Ask for price

Ranger DNA Polymerase

BIO-21121 250 Units Ask for price

Ranger DNA Polymerase

BIO-21121/S Sample Ask for price

Ranger DNA Polymerase

BIO-21122 500 Units Ask for price

Ready? DNA Polymerase

M1146-1000
EUR 398.4

Ready? DNA Polymerase

M1146-10000
EUR 1279.2

Ready? DNA Polymerase

M1146-5000
EUR 960

Breeze? DNA Polymerase

M1148-1000
EUR 1246.8

Breeze? DNA Polymerase

M1148-250
EUR 464.4

Distant? DNA Polymerase

M1150-1000
EUR 823.2

Distant? DNA Polymerase

M1150-250
EUR 333.6

Advance? DNA Polymerase

M1151-1000
EUR 895.2

Advance? DNA Polymerase

M1151-250
EUR 373.2

Outstretched? DNA Polymerase

M1152-1000
EUR 960

Outstretched? DNA Polymerase

M1152-250
EUR 398.4

T4 DNA Polymerase

M1211-100
EUR 385.2

Phi29 DNA Polymerase

M1239-1000
EUR 373.2

AceTaq DNA Polymerase

P401-d1 250 U
EUR 163.2

AceTaq DNA Polymerase

P401-d2 1000 U
EUR 272.4

AceTaq DNA Polymerase

P401-d3 3000 U
EUR 536.4

Taq DNA Polymerase

L7051001 1000 unit
EUR 234
Description: Premade ready to use kits will always come in handy. Get your experiment done right form the first try by using a validated kit with perfectly balanced reagents proportions and compatibility and by following a clear protocol.

Taq DNA Polymerase

L7051200 200 unit
EUR 104.4
Description: Premade ready to use kits will always come in handy. Get your experiment done right form the first try by using a validated kit with perfectly balanced reagents proportions and compatibility and by following a clear protocol.

T4 DNA Polymerase

N101-01 2000 U
EUR 278.4

Kodaq DNA Polymerase

G498 250 U (50 ul)
EUR 104.4

Kodaq DNA Polymerase

G499 1000 U (200 ul)
EUR 189.6

Pfu DNA Polymerase

S116 1x250 units
EUR 91.2

Pfu DNA Polymerase

S117 2x250 units
EUR 112.8

Pfu DNA Polymerase

S118 10x250 units
EUR 363.6

Bst DNA Polymerase

S600 2000 U
EUR 128.4

100 BP DNA LADDER, 500UL PER KIT

M-DNA-100BP 1/pk
EUR 87.6
Description: Bioscience Mol Bio; DNA Ladder

1 KB DNA LADDER, 500UL PER KIT

M-DNA-1KB 1/pk
EUR 84
Description: Bioscience Mol Bio; DNA Ladder

DNA Polymerase alpha Antibody

DF3082 200ul
EUR 420

DNA Polymerase beta Antibody

DF6552 200ul
EUR 420

DNA Polymerase theta Antibody

DF13563 100ul
EUR 420

DNA Polymerase alpha Antibody

AF5235 200ul
EUR 420

DNA Polymerase beta Antibody

AF0148 200ul
EUR 420

DNA Polymerase beta antibody

70R-50226 100 ul
EUR 292.8
Description: Purified Polyclonal DNA Polymerase beta antibody

DNA Polymerase lambda antibody

70R-32186 100 ug
EUR 392.4
Description: Rabbit polyclonal DNA Polymerase lambda antibody

DNA Polymerase zeta antibody

70R-33711 100 ug
EUR 392.4
Description: Rabbit polyclonal DNA Polymerase zeta antibody

DNA Polymerase zeta antibody

70R-33727 100 ug
EUR 392.4
Description: Rabbit polyclonal DNA Polymerase zeta antibody

DNA Polymerase theta antibody

70R-33737 100 ug
EUR 392.4
Description: Rabbit polyclonal DNA Polymerase theta antibody

DNA Polymerase beta antibody

70R-31161 100 ug
EUR 392.4
Description: Rabbit polyclonal DNA Polymerase beta antibody

DNA Polymerase alpha antibody

70R-31578 100 ug
EUR 392.4
Description: Rabbit polyclonal DNA Polymerase alpha antibody

DNA Polymerase beta Antibody

49605-100ul 100ul
EUR 399.6

DNA Polymerase beta Antibody

49605-50ul 50ul
EUR 286.8

DNA polymerase gamma antibody

70R-12532 100 ul
EUR 548.4
Description: Affinity purified Rabbit polyclonal DNA polymerase gamma antibody

Recombinant Pfu DNA Polymerase

7-03322 100U Ask for price

Recombinant Pfu DNA Polymerase

7-03323 500U Ask for price

Recombinant Pfu DNA Polymerase

7-03324 2500U Ask for price

Recombinant Taq DNA Polymerase

7-03634 1,000U Ask for price

Recombinant Taq DNA Polymerase

7-03635 3,000U Ask for price

Recombinant Taq DNA Polymerase

7-03636 10,000U Ask for price

DNA Polymerase gamma antibody

22870-100ul 100ul
EUR 468

DNA Polymerase ζ Antibody

33602-100ul 100ul
EUR 302.4

DNA Polymerase ζ Antibody

33602-50ul 50ul
EUR 224.4

DNA Polymerase θ Antibody

33611-100ul 100ul
EUR 302.4

DNA Polymerase θ Antibody

33611-50ul 50ul
EUR 224.4

DNA Polymerase α Antibody

33665-100ul 100ul
EUR 302.4

DNA Polymerase α Antibody

33665-50ul 50ul
EUR 224.4

DNA Polymerase beta Antibody

35404-100ul 100ul
EUR 468

DNA Polymerase λ Antibody

34098-100ul 100ul
EUR 302.4

DNA Polymerase λ Antibody

34098-50ul 50ul
EUR 224.4

DNA Polymerase β Antibody

33357-100ul 100ul
EUR 302.4

DNA Polymerase β Antibody

33357-50ul 50ul
EUR 224.4

Taq DNA polymerase (+dNTPs)

02-001 200U
EUR 183.6
Description: The Taq DNA polymerase (+dNTPs) is available in Europe and for worldwide shipping via Gentaur.

Taq DNA polymerase (+dNTPs)

02-001-5 5×200U
EUR 308.4
Description: The Taq DNA polymerase (+dNTPs) is available in Europe and for worldwide shipping via Gentaur.

Taq DNA polymerase (-dNTPs)

02-011 200U
EUR 168
Description: The Taq DNA polymerase (-dNTPs) is available in Europe and for worldwide shipping via Gentaur.

Taq DNA polymerase (-dNTPs)

02-011-5 5×200U
EUR 261.6
Description: The Taq DNA polymerase (-dNTPs) is available in Europe and for worldwide shipping via Gentaur.

Pfu DNA polymerase (+dNTPs)

02-021 200U
EUR 214.8
Description: The Pfu DNA polymerase (+dNTPs) is available in Europe and for worldwide shipping via Gentaur.

Pfu DNA polymerase (+dNTPs)

02-021-5 5×200U
EUR 402
Description: The Pfu DNA polymerase (+dNTPs) is available in Europe and for worldwide shipping via Gentaur.

Pfu DNA polymerase (-dNTPs)

02-031 200U
EUR 199.2
Description: The Pfu DNA polymerase (-dNTPs) is available in Europe and for worldwide shipping via Gentaur.

Pfu DNA polymerase (-dNTPs)

02-031-5 5×200U
EUR 339.6
Description: The Pfu DNA polymerase (-dNTPs) is available in Europe and for worldwide shipping via Gentaur.

DNA polymerase ? (C-His)

10-105 50ug
EUR 637.2
Description: The DNA polymerase ? (C-His) is available in Europe and for worldwide shipping via Gentaur.

T DNA Polymerase Enzyme

abx071008-250U 250 U
EUR 226.8

T DNA Polymerase Enzyme

abx071008-3kU 3 kU
EUR 543.6

T DNA Polymerase Enzyme

abx071008-500U 500 U
EUR 260.4

TopTaq DNA Polymerase Enzyme

abx071014-250U 250 U
EUR 276

TopTaq DNA Polymerase Enzyme

abx071014-3kU 3 kU
EUR 944.4

TopTaq DNA Polymerase Enzyme

abx071014-500U 500 U
EUR 360

EasyPfu DNA Polymerase Enzyme

abx071016-250U 250 U
EUR 226.8

EasyPfu DNA Polymerase Enzyme

abx071016-3kU 3 kU
EUR 526.8

EasyPfu DNA Polymerase Enzyme

abx071016-500U 500 U
EUR 260.4

FastPfu DNA Polymerase Enzyme

abx071018-250U 250 U
EUR 309.6

FastPfu DNA Polymerase Enzyme

abx071018-3kU 3 kU
EUR 1111.2

FastPfu DNA Polymerase Enzyme

abx071018-500U 500 U
EUR 376.8

Taq DNA Polymerase (Recombinant)

abx073521-10000U 10000 U
EUR 710.4

Taq DNA Polymerase (Recombinant)

abx073521-1000U 1000 U
EUR 276

Taq DNA Polymerase (Recombinant)

abx073521-3000U 3000 U
EUR 393.6

Pfu DNA Polymerase (Recombinant)

abx074017-100U 100 U
EUR 393.6

Pfu DNA Polymerase (Recombinant)

abx074017-2500U 2500 U
EUR 1528.8

Pfu DNA Polymerase (Recombinant)

abx074017-500U 500 U
EUR 526.8

DNA Polymerase beta Antibody

20-abx013060
  • EUR 376.80
  • EUR 117.60
  • EUR 477.60
  • EUR 594.00
  • 100 ug
  • 10 ug
  • 200 ug
  • 300 µg

DNA Polymerase alpha Antibody

20-abx013375
  • EUR 376.80
  • EUR 117.60
  • EUR 477.60
  • EUR 594.00
  • 100 ug
  • 10 ug
  • 200 ug
  • 300 µg

DNA Polymerase beta Antibody

20-abx008852
  • EUR 360.00
  • EUR 526.80
  • EUR 226.80
  • 100 ul
  • 200 ul
  • 30 ul

DNA Polymerase beta Antibody

ABF0148 100 ug
EUR 525.6

DNA polymerase beta Antibody

abx232432-100ug 100 ug
EUR 577.2

Taq DNA Polymerase (500U)

9K-001-0001 500U
EUR 209.29

Taq DNA Polymerase (3000U)

9K-001-0002 3000U
EUR 582

Taq DNA Polymerase (6000U)

9K-001-0033 6000U
EUR 1234.5

Taq DNA Polymerase (1000U)

9K-001-0035 1000U
EUR 418.62

MyTaq Red DNA Polymerase

BIO-21108 500 units Ask for price

MyTaq Red DNA Polymerase

BIO-21108/S Sample Ask for price

MyTaq Red DNA Polymerase

BIO-21109 2500 units Ask for price

MyTaq Red DNA Polymerase

BIO-21110 5000 units Ask for price

MyTaq HS DNA Polymerase

BIO-21111 250 units Ask for price

MyTaq HS DNA Polymerase

BIO-21111/S Sample Ask for price

MyTaq HS DNA Polymerase

BIO-21112 1000 units Ask for price

MyTaq HS DNA Polymerase

BIO-21113 2500 units Ask for price

Robust Ready? DNA Polymerase

M1147-1000
EUR 1149.6

Robust Ready? DNA Polymerase

M1147-250
EUR 438

Fire Start? DNA Polymerase

M1149-1000
EUR 764.4

Fire Start? DNA Polymerase

M1149-250
EUR 307.2

Blood Advance? DNA Polymerase

M1153-100
EUR 268.8

Blood Advance? DNA Polymerase

M1153-400
EUR 568.8

Taq Plus DNA Polymerase

P201-01 250 U
EUR 147.6

Taq Plus DNA Polymerase

P201-02 1000 U
EUR 213.6

Taq Plus DNA Polymerase

P201-03 3000 U
EUR 382.8

amfiSure CloneEasy DNA Polymerase

P0325-010 100 units
EUR 278.4

amfiSure CloneEasy DNA Polymerase

P0325-025 250 units
EUR 481.2

Champagne Taq DNA Polymerase

P122-d1 500 U (2.5 U/μl)
EUR 212.4

Champagne Taq DNA Polymerase

P122-d2 500 U (5 U/μl)
EUR 212.4

Champagne Taq DNA Polymerase

P122-d3 500 U (10 U/μl)
EUR 212.4

Hotstart Taq DNA Polymerase

L7052001 1000 units
EUR 681.6
Description: Premade ready to use kits will always come in handy. Get your experiment done right form the first try by using a validated kit with perfectly balanced reagents proportions and compatibility and by following a clear protocol.

Hotstart Taq DNA Polymerase

L7052250 250 units
EUR 234
Description: Premade ready to use kits will always come in handy. Get your experiment done right form the first try by using a validated kit with perfectly balanced reagents proportions and compatibility and by following a clear protocol.

HiFi DNA Polymerase wiodNTP

FYT033-500U 500U Ask for price

Accu DNA Polymerase wiodNTPs

FYT044-500U 500U Ask for price

Taq DNA Polymerase (PCR)

G008 5000 U (1.0 ml)
EUR 246

Taq DNA Polymerase (PCR)

G009 1000 U (200 ul)
EUR 104.4

HotStart DNA Polymerase (PCR)

G011 250 U (50 ul)
EUR 98.4

HotStart DNA Polymerase (PCR)

G039 1000 U (200 ul)
EUR 189.6

Taq DNA Polymerase (PCR)

G126 10000 U (2 x 1.0 ml)
EUR 370.8

TaqFast DNA Polymerase (PCR)

G277 250 U (50 ul)
EUR 87.6

TaqFast DNA Polymerase (PCR)

G278 1000 U (200 ul)
EUR 160.8

Bloodirect DNA Polymerase (PCR)

G462 100 U (100 ul)
EUR 73.2

Bloodirect DNA Polymerase (PCR)

G463 400 U (400 ul)
EUR 104.4

SensTaq HotStart DNA Polymerase

G937 250U
EUR 104.4

SensTaq HotStart DNA Polymerase

G938 1000U
EUR 189.6

Maximo Taq DNA Polymerase

S101 500 units
EUR 55.2

Maximo Taq DNA Polymerase

S102 5x500 units
EUR 128.4

Maximo Taq DNA Polymerase

S103 20x500 units
EUR 393.6


In order to develop a test to identify the genes responsible for antibiotic resistance in gram-negative bacteria, it has been found that the standard of Taq DNA polymerase (not DNA-free) blaTEM contaminated with fragments of genes that varied in length and number. BlaTEM complete gene sequences are absent or detected in very small amounts. We developed an approach to avoid false positive findings caused by contamination in the polymerase gene sequence blaTEM conventional.