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
BA00105 2500U
EUR 278.4
Description: High quality Taq 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.

Taq DNA polymerase
BT10101 500U
EUR 96
Description: High quality Taq polymerase for different PCR variations and downstream applications.

Taq DNA polymerase
BT10102 1000U
EUR 130.8
Description: High quality Taq polymerase for different PCR variations and downstream applications.

Taq DNA polymerase
BT10103 2000U
EUR 200.4
Description: High quality Taq polymerase for different PCR variations and downstream applications.

Taq DNA Polymerase
9001-2500 each
EUR 614.4

Taq DNA Polymerase
9001-500 each
EUR 229.2

PFU DNA Polymerase
9003-2500 each
EUR 614.4

PFU DNA Polymerase
9003-500 each
EUR 216

Laq? DNA Polymerase
9004-2500 each
EUR 732

Laq? DNA Polymerase
9004-500 each
EUR 261.6

Taq DNA Polymerase
L7051001 1000 unit
EUR 159

Taq DNA Polymerase
L7051200 200 unit
EUR 41

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

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

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 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

Ready? DNA Polymerase
M1146-1000 each
EUR 398.4

Ready? DNA Polymerase
M1146-10000 each
EUR 1279.2

Ready? DNA Polymerase
M1146-5000 each
EUR 960

Phi29 DNA Polymerase
M1239-1000 each
EUR 373.2

Kodaq DNA Polymerase
G498 250 U (50 ul)
EUR 104.4

Kodaq DNA Polymerase
G499 1000 U (200 ul)
EUR 189.6

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.

BIOTAQ DNA Polymerase
BIO-21040 500 Units Ask for price

BIOTAQ DNA Polymerase
BIO-21060 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

HotTaq DNA polymerase
BT10201 500U
EUR 144
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.

HotTaq DNA polymerase
BT10202 1000U
EUR 228
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.

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

Breeze? DNA Polymerase
M1148-1000 each
EUR 1246.8

Breeze? DNA Polymerase
M1148-250 each
EUR 464.4

AceTaq DNA Polymerase
P401-MD1 250 U
EUR 46.4

Taq DNA Polymerase (500U)
9K-001-0001 500U
EUR 209.29

Distant? DNA Polymerase
M1150-1000 each
EUR 823.2

Distant? DNA Polymerase
M1150-250 each
EUR 333.6

Advance? DNA Polymerase
M1151-1000 each
EUR 895.2

Advance? DNA Polymerase
M1151-250 each
EUR 373.2

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

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

Taq HS DNA Polymerase
P132-d1 500 U
EUR 38.28

Taq HS DNA Polymerase
P132-d2 1,000 U
EUR 71.34

Taq HS DNA Polymerase
P132-d3 5,000 U
EUR 324.8

Outstretched? DNA Polymerase
M1152-1000 each
EUR 960

Outstretched? DNA Polymerase
M1152-250 each
EUR 398.4

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.

Taq DNA Polymerase (PCR)
G008 5000 U (1.0 ml)
EUR 246

Taq DNA Polymerase (PCR)
G009 1000 U (200 ul)
EUR 104.4

Taq DNA Polymerase (PCR)
G126 10000 U (2 x 1.0 ml)
EUR 370.8

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

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

ACTaq? Taq DNA Polymerase, 1000U
E2100-1000U each
EUR 260.4

T4 DNA Polymerase - 500 units
3222 1/EA
EUR 247.2

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.

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

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

ACTaq? Long DNA Polymerase, 1000U
E2200-1000U each
EUR 282

Taq DNA Polymerase - 5,000 units
3245 1/EA
EUR 620.4

Fire Start? DNA Polymerase
M1149-1000 each
EUR 764.4

Fire Start? DNA Polymerase
M1149-250 each
EUR 307.2

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

Taq DNA Polymerase, 5units/㎕
T2200-050 500units
EUR 157.2

Taq DNA Polymerase, 5units/㎕
T2200-100 2x500 units
EUR 230.4

Taq DNA Polymerase, 5units/㎕
T2200-200 4x500 units
EUR 375.6

ACTaq? Taq DNA Polymerase, 250U
E2100-250U each
EUR 122.4

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

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

TaqFast DNA Polymerase (PCR)
G277 250 U (50 ul)
EUR 87.6

TaqFast DNA Polymerase (PCR)
G278 1000 U (200 ul)
EUR 160.8

Fast-Taq DNA Polymerase (500U)
9K-001-0036 500U
EUR 420.65

ACTaq? Long DNA Polymerase, 250U
E2200-250U each
EUR 127.2

Hotstart Taq DNA Polymerase
L7052001 1000 units
EUR 567

Hotstart Taq DNA Polymerase
L7052250 250 units
EUR 159

Robust Ready? DNA Polymerase
M1147-1000 each
EUR 1149.6

Robust Ready? DNA Polymerase
M1147-250 each
EUR 438

HotStart DNA Polymerase (PCR)
G011 250 U (50 ul)
EUR 98.4

HotStart DNA Polymerase (PCR)
G039 1000 U (200 ul)
EUR 189.6

HiFi DNA Polymerase wiodNTP
FYT033-500U 500U Ask for price

T4 DNA Polymerase, 5u/ul
BEP0061 100U
EUR 153.97

T4 DNA Polymerase (5,000 U/ml)
RK20539 25 U Ask for price

DNA polymerase beta Antibody
abx232432-100ug 100 ug
EUR 577.2

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

DNA Polymerase beta Antibody
AF0148 200ul
EUR 420

DNA Polymerase beta Antibody
49605-100ul 100ul
EUR 399.6

DNA Polymerase beta Antibody
49605-50ul 50ul
EUR 286.8

DNA Polymerase beta antibody
70R-31161 100 ug
EUR 392.4
Description: Rabbit polyclonal DNA Polymerase beta 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 beta antibody
70R-50226 100 ul
EUR 292.8
Description: Purified Polyclonal DNA Polymerase beta antibody

DNA Polymerase beta Antibody
35404-100ul 100ul
EUR 468

DNA Polymerase beta Antibody
ABF0148 100 ug
EUR 525.6

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

DNA Polymerase beta Antibody
DF6552 200ul
EUR 420

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

Blood Advance? DNA Polymerase
M1153-100 each
EUR 268.8

Blood Advance? DNA Polymerase
M1153-400 each
EUR 568.8

Accu DNA Polymerase wiodNTPs
FYT044-500U 500U Ask for price

Taq DNA Polymerase, 5u/ul
B0089(D0089) 5X200U
EUR 112.2

Pfu DNA Polymerase, 5u/ul
B0093(D0090P) 500U
EUR 164.4

DNA Polymerase I, 10u/ul
BEP0041 500U
EUR 128.9

Tsg DNA Polymerase, 5u/ul
D0081 5X200U
EUR 112.2

DNA Polymerase gamma antibody
22870-100ul 100ul
EUR 468

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

DNA Polymerase alpha Antibody
AF5235 200ul
EUR 420

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

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

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

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 alpha Antibody
DF3082 200ul
EUR 420

DNA Polymerase theta Antibody
DF13563 100ul
EUR 420

Bloodirect DNA Polymerase (PCR)
G462 100 U (100 ul)
EUR 73.2


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.