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

9001-2500 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

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.

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

EQ005 100μl/500U
EUR 50

Pfu DNA Polymerase

EQ008 50μl
EUR 68

Pfu DNA Polymerase

enz-265 100U
EUR 145
Description: Recombinant Pfu DNA Polymerase

Taq DNA Polymerase

G009 1000 U (200 ul)
EUR 20
Description: Taq DNA Polymerase is a highly thermostable DNA Polymerase that catalyzes the 5’-3’ synthesis of DNA. This polymerase has 5’-3’ exonuclease activities, lacks 3’-5’ exonuclease activity, and produces 3’-dA-tailed amplicons. PCR products made with Taq can be used with TA cloning vectors.

Taq DNA Polymerase

L7051001 1000 unit
EUR 167

Taq DNA Polymerase

L7051200 200 unit
EUR 43

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

Taq DNA Polymerase

MB101-0500 500 U/100 μl
EUR 10

Taq DNA Polymerase

abx299725-100g 100 µg Ask for price

Taq DNA Polymerase

abx299725-20g 20 µg
EUR 337.5

Taq DNA Polymerase

abx299725-50g 50 µg Ask for price

DNA Polymerase Mix

abx461002-1096tests 10 × 96 tests Ask for price

DNA Polymerase Mix

abx461002-596tests 5 × 96 tests Ask for price

DNA Polymerase Mix

abx461002-96tests 96 tests
EUR 1262.5

Taq DNA Polymerase

abx461029-1096tests 10 × 96 tests
EUR 1787.5

Taq DNA Polymerase

abx461029-596tests 5 × 96 tests
EUR 1062.5

Taq DNA Polymerase

abx461029-96tests 96 tests
EUR 362.5

DNA Polymerase beta

E8ET1705-12 100ul
EUR 275
Description: Available in various conjugation types.

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

DNA polymerase delta

E8ER1907-56 100ul
EUR 275
Description: Available in various conjugation types.

Phi29 DNA Polymerase

E014 1000 U (100 μl)
EUR 145
Description: Phi29 DNA Polymerase is a highly processive polymerase which exhibits a strong strand-displacement function. These functions allow for highly efficient isothermal amplification of circular or linear DNA templates via rolling circle amplification (RCA), multiple displacement amplification (MDA) and/or whole genome amplification (WGA). Phi29 DNA Polymerase has extremely high fidelity due to its inherent 3’→5’ exonuclease activity and can amplify from very small amounts of starting templates.

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

Kodaq DNA Polymerase

G498 250 U (50 ul)
EUR 104.4

Kodaq DNA Polymerase

G499 1000 U (200 ul)
EUR 189.6

Phi29 DNA Polymerase

PCR-381L 5 x 200units
EUR 255
Description: Bacillus subtilis phage phi29, recombinant, E. coli

Phi29 DNA Polymerase

PCR-381S 200units
EUR 63.8
Description: Bacillus subtilis phage phi29, recombinant, E. coli

Ultra DNA Polymerase

PCR-391L 5 x 100units
EUR 486.7
Description: also known as Phusion High-Fidelity DNA Polymerase

Ultra DNA Polymerase

PCR-391S 100units
EUR 121.7
Description: also known as Phusion High-Fidelity DNA Polymerase

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

phi29 DNA Polymerase

RK21002 1250U
EUR 4

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

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.

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.

Breeze? DNA Polymerase

M1148-1000 each
EUR 1246.8

Breeze? DNA Polymerase

M1148-250 each
EUR 464.4

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

AceTaq DNA Polymerase

P401-MD1 250 U
EUR 58

Taq DNA Polymerase (500U)

9K-001-0001 500U
EUR 209.29

BlasTaq DNA Polymerase

E36G894 400 rxn (200 μl)
EUR 528.57

BlasTaq DNA Polymerase

G894 400 rxn (200 μl)
EUR 80
Description: BlasTaq™ DNA Polymerase is a strategically-engineered, next generation Taq Polymerasethat has rapid extension rates and robust performance. With specialized reactionconditions, this polymerase provides increased processivity, yields, and sensitivity, whileshortening reaction times by up to 70%, compared to wild-type Taq DNA polymerase.BlasTaq™ has 5’-3’ polymerase and 5’-3’ exonuclease activities, lacks 3’-5’ exonucleaseactivity, and produces 3’-dA-tailed amplicons. PCR products made with BlasTaq™ can beused with TA cloning vectors.

BlasTaq DNA Polymerase

G894-S 200 rxn (100 ul)
EUR 45
Description: BlasTaq™ DNA Polymerase is a strategically-engineered, next generation Taq Polymerasethat has rapid extension rates and robust performance. With specialized reactionconditions, this polymerase provides increased processivity, yields, and sensitivity, whileshortening reaction times by up to 70%, compared to wild-type Taq DNA polymerase.BlasTaq™ has 5’-3’ polymerase and 5’-3’ exonuclease activities, lacks 3’-5’ exonucleaseactivity, and produces 3’-dA-tailed amplicons. PCR products made with BlasTaq™ can beused with TA cloning vectors.

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

DNA Polymerase Antibody

8C10144 50ug
EUR 368
Description: DNA Polymerase ζ Antibody

DNA Polymerase Antibody

E11-11836C 100μg
EUR 225
Description: Available in various conjugation types.

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 47.85

Taq HS DNA Polymerase

P132-d2 1,000 U
EUR 89.18

Taq HS DNA Polymerase

P132-d3 5,000 U
EUR 406

TM Taq DNA Polymerase

TBS4010 1000 U
EUR 60

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)

G126 10000 U (2 x 1.0 ml)
EUR 370.8

Taq DNA Polymerase Mix

abx461043-1096tests 10 × 96 tests Ask for price

Taq DNA Polymerase Mix

abx461043-596tests 5 × 96 tests
EUR 1062.5

Taq DNA Polymerase Mix

abx461043-96tests 96 tests
EUR 337.5

Taq DNA Polymerase -1000 units

3243 1000units
EUR 115
Description: Intact Genomics Taq DNA Polymerase is a thermostable DNA polymerase that possesses a 5´→3´ polymerase activity (1, 2) and a 5´ flap endonuclease activity (3, 4). This product is supplied with 10x PCR reaction buffer, containing MgCl2, which produces a final Mg2+ concentration of 1.5 mM. Ideal for primary extension reaction with DNA fragments having dA overhang on 3’ ends.Product Includes:Taq DNA Polymerase10x PCR Buffer with Mg2+5x Magic Enhancer

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

Long Amp DNA polymerase

MBT069-1000U 1 unit
EUR 150.52
Description: Long Amp DNA polymerase

Long Amp DNA polymerase

MBT069-100U 1 unit
EUR 22.01
Description: Long Amp DNA polymerase

Long Amp DNA polymerase

MBT069-200U 1 unit
EUR 40.48
Description: Long Amp DNA polymerase

Long Amp DNA polymerase

MBT069-500U 1 unit
EUR 88.98
Description: Long Amp DNA polymerase

ACTaq? Taq DNA Polymerase, 1000U

E2100-1000U each
EUR 260.4

T4 DNA Polymerase - 500 units

3222 1/EA
EUR 175
Description: Intact Genomics T4 DNA Polymerase has both a DNA-dependent DNA polymerase activity and a potent 3´→5´ exonuclease activity. Product Includes:T4 DNA Polymerase10x T4 DNA Polymerase Buffer

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

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 delta p50

E8ER1907-57 100ul
EUR 275
Description: Available in various conjugation types.

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

Phi29 MAX DNA Polymerase

N106-01 250 U
EUR 24

Phi29 MAX DNA Polymerase

N106-02 1,250 U
EUR 95.5

Hi-Temp DNA polymerase

MBT070-100U 1 unit
EUR 105.46
Description: Hi-Temp DNA polymerase

Hi-Temp DNA polymerase

MBT070-200U 1 unit
EUR 200.37
Description: Hi-Temp DNA polymerase

Hi-Temp DNA polymerase

MBT070-500U 1 unit
EUR 486.17
Description: Hi-Temp DNA polymerase

T DNA Polymerase Enzyme

abx071008-100g 100 g
EUR 275

T DNA Polymerase Enzyme

abx071008-25g 25 g
EUR 237.5

ACTaq? Long DNA Polymerase, 1000U

E2200-1000U each
EUR 282

Taq DNA Polymerase - 5,000 units

3245 1/EA
EUR 449
Description: Intact Genomics Taq DNA Polymerase is a thermostable DNA polymerase that possesses a 5´→3´ polymerase activity (1, 2) and a 5´ flap endonuclease activity (3, 4). This product is supplied with 10x PCR reaction buffer, containing MgCl2, which produces a final Mg2+ concentration of 1.5 mM. Ideal for primary extension reaction with DNA fragments having dA overhang on 3’ ends.Product Includes:Taq DNA Polymerase10x PCR Buffer with Mg2+5x Magic Enhancer

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

Hi-Proof DNA Polymerase

MBT068-100U 1 unit
EUR 42.75
Description: Hi-Proof DNA Polymerase

Hi-Proof DNA Polymerase

MBT068-200U 1 unit
EUR 74.11
Description: Hi-Proof DNA Polymerase

Hi-Proof DNA Polymerase

MBT068-500U 1 unit
EUR 120.57
Description: Hi-Proof DNA Polymerase

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

Rabbit Polyclonal antibody to DNA polymerase gamma (polymerase (DNA directed), gamma)

TA308753 100 µl Ask for price

DNA Polymerase β Antibody

8C0173 50ug
EUR 368
Description: DNA Polymerase β Antibody

DNA Polymerase θ Antibody

8C10173 50ug
EUR 368
Description: DNA Polymerase θ Antibody

DNA Polymerase α Antibody

8C10322 50ug
EUR 368
Description: DNA Polymerase α Antibody

DNA Polymerase λ Antibody

8C11836 50ug
EUR 368
Description: DNA Polymerase λ Antibody

DNA Polymerase β Antibody

33357 100ul
EUR 319

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
EUR 319

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
EUR 319

DNA Polymerase θ Antibody

33611-100ul 100ul
EUR 302.4

DNA Polymerase θ Antibody

33611-50ul 50ul
EUR 224.4


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.