Directed co-evolution of interacting protein-peptide pairs by compartmentalized two-hybrid replication (C2HR)

Directed co-evolution of interacting protein-peptide pairs by compartmentalized two-hybrid replication (C2HR)

The methodologies of the directed evolution benefit from quantitative reading quantitatively genotype at the phenotype. We have therefore designed a method flowing from protein-peptide interactions to dynamic reading provided by a modified polymerase DNA. The melting of a processivity clamping protein on a polymerase thermostable nucleic acid allows a polymerase activity and an amplification of the DNA in high prohibition salt buffers. Here, we recap this phenotype by indirectly coupling the SSO7D processing clamp to the TAQ DNA polymerase via a respective fusion to a pair of high affinity peptide proteins and thermostable interaction. Escherichia coli cells co-expressing peptide-peptide-peptide pairs can be used directly in polymerase chain reactions to determine the relative interaction resistors by measuring amplic yields. The conditional activity of the polymerase is also used to connect the genotype to the phenotype of the interactive peptide pairs co-expressed in E. coli using the phased evolution platform for the compartmentalized automatic replication.

We validate this approach, called the replication with two hybrid compartmentalized, by selecting for the high affinity peptides that bind two models of model proteins: Spycacher and the large fragment of Nanoluc Luciferase. We demonstrate more co-evolution directed by hiking both protein and peptide components of the SpycCatcher-Spytag pair and by co-selecting for functional interactive variants. An important toxin-antitoxin (TA) HOK / Sok system, encoded by the plasmid R1 of Escherichia coli, is involved in the wake of post-segregation cells that have lost the plasmid. The deadly properties of HOK protein have been used for environmental containment of microbes and the development of potential vaccine candidates. We aim to demonstrate the powerful antimicrobial property of 19 fragment of the HOK peptide amino acid (AA) terminal.

This has been accomplished by designing a conditional suicide system based on HOK gene expression, cloned in an anhydroteracyclacin (ATC) inducible vector – PASK75. Thermal shocks and electroporation were used for the transformation of Escherichia coli cells and Vibrio Cholerae respectively. The minimum induction concentration (medium C) of the ATC, determined by analyzing the expression of the green fluorescent protein cloned separately in the PASK75 vector, was 30 ng / ml. As the HOK gene has been synthesized by Novo (using the reaction of the recombinant polymerase chain) in our study, various random size HOK fragments were generated (as a result of the possible nature of Taq. polymerase).

 

Building better DNA polymerases: Engineering Replication of Expanded Genetic Alphabets

 

DNA polymerases are now used in all scientific research, biotechnology and medicine, in part for their ability to interact with non-natural forms of DNA created by synthetic biologists. Here, especially, natural DNA polymerases often do not have “performance specifications” needed for processing technologies. This creates a need for rational (or semi-rational) scientific engineering engineering in order to identify variants that reproduce non-natural basic pairs (UBP), non-natural backbones, tags or other features evolutionually new from the Non-natural DNA. In this review, we provide a brief overview of the chemistry and properties of the replica DNA polymerases and their evolved variants, with a focus on the Klenow fragment of Taq DNA polymerase (Klentaq).

We describe studies of structural, enzymatic and comparative molecular dynamics of wild-type variants and klentaq, complex with natural or non-canonic substrates. The combination of these methods makes it possible to understand how the specific amino acid substitutions remote from the active site in a variant of the Klentaq DNA polymerase (ZP Klentaq) contribute to its ability to reproduce non-natural basic pairs (UBP). With improved efficiency compared to Klentaq. This approach can therefore be used to guide any future rational engineering of replicative DNA polymerases.

 Directed co-evolution of interacting protein-peptide pairs by compartmentalized two-hybrid replication (C2HR)
Directed co-evolution of interacting protein-peptide pairs by compartmentalized two-hybrid replication (C2HR)

Evaluation of the comparability of different methods of DNA extraction and amplification in microbial community profiling of intestines

 

Automated and broadband technologies are becoming increasingly common in microbiome studies to reduce costs and increase efficiency. However, in microbiome studies, small differences in methodology – including storage conditions, wet laboratory methods, sequencing platforms and data analysis – can affect the reproducibility and comparability of data between the studies.

The effects of broadband methods have been limited, including microfluidic PCR technologies. In this article, we compare two extraction methods (the mini Qiaamp DNA stool kit and the Mobio PowerSoil DNA insulation kit), two Taq polymerase enzymes (Mytaq HS Red Mix and Lock II PCR Tougymix) , two sets of primers (V3-V4 and V4 -V5) and two amplification methods (a two-step PCR protocol and a preparation of the amplicon library on the fluidigm access network system that allows multiplexing automated primers). The profiles of the Gut microbial community have been considerably affected by all variables. Although there has been no significant difference in alpha diversity measured between the two extraction methods, there was an effect of the extraction method on the community composition measured by unrestricted unifruc distances. The amplification method and primers have had a significant effect on alpha diversity and community composition.

DNA Polymerase ζ Conjugated Antibody

C33602 100ul
EUR 397

Anti-Taq DNA Polymerase Antibody

48369-100ul 100ul
EUR 333

Anti-Taq DNA Polymerase Antibody

48369-50ul 50ul
EUR 239

Anti-DNA Polymerase thet Antibody

A02205 100ul
EUR 397
Description: Rabbit Polyclonal DNA Polymerase thet Antibody. Validated in IHC and tested in Human.

anti- DNA polymerase beta antibody

FNab02432 100µg
EUR 505.25
  • Immunogen: polymerase(DNA directed), beta
  • Uniprot ID: P06746
  • Research Area: Metabolism
Description: Antibody raised against DNA polymerase beta

Anti-DNA polymerase beta antibody

PAab02432 100 ug
EUR 355

Anti-DNA polymerase theta antibody

STJ71548 100 µg
EUR 260

Anti-DNA polymerase eta Antibody

STJ502747 100 µg
EUR 476

anti-DNA Polymerase epsilon

YF-PA19269 50 ul
EUR 363
Description: Mouse polyclonal to DNA Polymerase epsilon

anti-DNA Polymerase epsilon

YF-PA19270 50 ul
EUR 363
Description: Mouse polyclonal to DNA Polymerase epsilon

anti-DNA Polymerase epsilon

YF-PA19271 50 ug
EUR 363
Description: Mouse polyclonal to DNA Polymerase epsilon

anti-DNA Polymerase epsilon

YF-PA19272 100 ug
EUR 403
Description: Rabbit polyclonal to DNA Polymerase epsilon

anti-DNA polymerase beta

YF-PA13864 50 ug
EUR 363
Description: Mouse polyclonal to DNA polymerase beta

anti-DNA polymerase beta

YF-PA13865 100 ul
EUR 403
Description: Rabbit polyclonal to DNA polymerase beta

anti-DNA polymerase eta

YF-PA13871 50 ug
EUR 363
Description: Mouse polyclonal to DNA polymerase eta

anti-DNA polymerase eta

YF-PA13872 100 ug
EUR 403
Description: Rabbit polyclonal to DNA polymerase eta

anti-DNA polymerase alpha

YF-PA24412 50 ul
EUR 334
Description: Mouse polyclonal to DNA polymerase alpha

anti-DNA polymerase beta

YF-PA24413 50 ul
EUR 334
Description: Mouse polyclonal to DNA polymerase beta

anti-DNA Polymerase epsilon

YF-PA26240 50 ul
EUR 334
Description: Mouse polyclonal to DNA Polymerase epsilon

anti-DNA polymerase? antibody (rabbit antiserum)

70-041 50uL
EUR 414
Description: The anti-DNA polymerase? antibody (rabbit antiserum) is available in Europe and for worldwide shipping via Gentaur.

Anti-DNA Polymerase lambda/POLL Antibody

A00959-2 100ug/vial
EUR 294

Anti-DNA Polymerase iota/POLI Antibody

A01376-1 100ug/vial
EUR 294

Anti-POLK/Dna Polymerase Kappa Antibody

A01987 100ug
EUR 432
Description: Goat Polyclonal POLK/Dna Polymerase Kappa Antibody. Validated in WB and tested in Human.

Anti-Taq DNA Polymerase Conjugated Antibody

C48369 100ul
EUR 397

Anti-DNA Polymerase beta Monoclonal Antibody

M01946 100ug
EUR 397
Description: Rabbit Monoclonal DNA Polymerase beta Antibody. Validated in IP, IF, WB and tested in Human, Mouse, Rat.

Anti-DNA Polymerase beta/POLB Antibody

PB9812 100ug/vial
EUR 294

Anti-DNA polymerase eta/POLH Antibody

PB9813 100ug/vial
EUR 294

Anti-DNA polymerase eta Antibody BIOTIN

STJ502748 100 µg
EUR 586

Anti-DNA polymerase eta Antibody FITC

STJ502749 100 µg
EUR 586

DNA Polymerase

ABD3082 100 ug
EUR 438

DNA Polymerase

ABF5235 100 ug
EUR 438

Anti-DNA Polymerase iota (8G9)

YF-MA11349 100 ug
EUR 363
Description: Mouse monoclonal to DNA Polymerase iota

Anti-DNA polymerase alpha (3C11)

YF-MA14797 100 ug
EUR 363
Description: Mouse monoclonal to DNA polymerase alpha

Anti-DNA Polymerase theta (1C11)

YF-MA17459 100 ug
EUR 363
Description: Mouse monoclonal to DNA Polymerase theta

anti-DNA Polymerase Kappa/POLK

YF-PA19042 50 ul
EUR 363
Description: Mouse polyclonal to DNA Polymerase Kappa/POLK

anti-DNA Polymerase Kappa/POLK

YF-PA19043 50 ug
EUR 363
Description: Mouse polyclonal to DNA Polymerase Kappa/POLK

anti-DNA Polymerase Kappa/POLK

YF-PA19044 100 ug
EUR 403
Description: Rabbit polyclonal to DNA Polymerase Kappa/POLK

anti-DNA polymerase delta p50

YF-PA13866 50 ug
EUR 363
Description: Mouse polyclonal to DNA polymerase delta p50

anti-DNA polymerase delta p50

YF-PA13867 100 ug
EUR 403
Description: Rabbit polyclonal to DNA polymerase delta p50

anti-DNA Polymerase epsilon p59

YF-PA24414 50 ul
EUR 334
Description: Mouse polyclonal to DNA Polymerase epsilon p59

anti-DNA Polymerase Kappa/POLK

YF-PA26181 50 ul
EUR 334
Description: Mouse polyclonal to DNA Polymerase Kappa/POLK

DNA Polymerase gamma antibody

22870-100ul 100ul
EUR 390

DNA polymerase gamma antibody

70R-12532 100 ul
EUR 457
Description: Affinity purified Rabbit polyclonal DNA polymerase gamma antibody

DNA Polymerase beta antibody

70R-31161 100 ug
EUR 327
Description: Rabbit polyclonal DNA Polymerase beta antibody

DNA Polymerase alpha antibody

70R-31578 100 ug
EUR 327
Description: Rabbit polyclonal DNA Polymerase alpha antibody

DNA Polymerase lambda antibody

70R-32186 100 ug
EUR 327
Description: Rabbit polyclonal DNA Polymerase lambda antibody

DNA Polymerase θ Antibody

33611-100ul 100ul
EUR 252

DNA Polymerase θ Antibody

33611-50ul 50ul
EUR 187

DNA Polymerase α Antibody

33665-100ul 100ul
EUR 252

DNA Polymerase α Antibody

33665-50ul 50ul
EUR 187

DNA Polymerase λ Antibody

34098-100ul 100ul
EUR 252

DNA Polymerase λ Antibody

34098-50ul 50ul
EUR 187

DNA Polymerase beta Antibody

35404-100ul 100ul
EUR 390

DNA Polymerase β Antibody

33357-100ul 100ul
EUR 252

DNA Polymerase β Antibody

33357-50ul 50ul
EUR 187

DNA Polymerase beta Antibody

49605-100ul 100ul
EUR 333

DNA Polymerase beta Antibody

49605-50ul 50ul
EUR 239

DNA Polymerase alpha Antibody

DF3082 200ul
EUR 304
Description: DNA Polymerase α Antibody detects endogenous levels of total DNA Polymerase α.

DNA Polymerase beta antibody

70R-50226 100 ul
EUR 244
Description: Purified Polyclonal DNA Polymerase beta antibody

DNA Polymerase zeta antibody

70R-33711 100 ug
EUR 327
Description: Rabbit polyclonal DNA Polymerase zeta antibody

DNA Polymerase zeta antibody

70R-33727 100 ug
EUR 327
Description: Rabbit polyclonal DNA Polymerase zeta antibody

DNA Polymerase theta antibody

70R-33737 100 ug
EUR 327
Description: Rabbit polyclonal DNA Polymerase theta antibody

DNA Polymerase beta Antibody

20-abx008852
  • EUR 300.00
  • EUR 439.00
  • EUR 189.00
  • 100 ul
  • 200 ul
  • 30 ul
  • Shipped within 5-10 working days.

DNA Polymerase beta Antibody

20-abx013060
  • EUR 314.00
  • EUR 98.00
  • EUR 398.00
  • EUR 495.00
  • 100 ug
  • 10 ug
  • 200 ug
  • 300 µg
  • Shipped within 5-10 working days.

DNA Polymerase alpha Antibody

20-abx013375
  • EUR 314.00
  • EUR 98.00
  • EUR 398.00
  • EUR 495.00
  • 100 ug
  • 10 ug
  • 200 ug
  • 300 µg
  • Shipped within 5-10 working days.

DNA Polymerase beta Antibody

AF0148 200ul
EUR 304
Description: DNA Polymerase beta antibody detects endogenous levels of total DNA Polymerase beta.

DNA Polymerase alpha Antibody

AF5235 200ul
EUR 304
Description: DNA Polymerase α Antibody detects endogenous levels of total DNA Polymerase α.

DNA polymerase beta Antibody

abx232432-100ug 100 ug
EUR 481
  • Shipped within 5-12 working days.

DNA Polymerase beta Antibody

ABF0148 100 ug
EUR 438

Rabbit Polyclonal antibody Anti-CRBN

Anti-CRBN 50 µg
EUR 349

anti- CD3 ζ(Phospho-Tyr142) antibody

FNab09792 100µg
EUR 548.75
  • Recommended dilution: WB: 1:500-1:2000
  • Immunogen: A synthesized peptide derived from human CD3 ζ(Phospho-Tyr142)
  • Uniprot ID: P20963
  • Research Area: Signal Transduction, Metabolism, Cardiovascular, Immunology, Cancer, Neuroscience, Stem Cells
Description: Antibody raised against CD3 ζ(Phospho-Tyr142)

Anti-DNA Polymerase Kappa/POLK (6F2)

YF-MA11510 100 ug
EUR 363
Description: Mouse monoclonal to DNA Polymerase Kappa/POLK

Anti-DNA Polymerase epsilon p59 (1A3)

YF-MA14798 100 ug
EUR 363
Description: Mouse monoclonal to DNA Polymerase epsilon p59

Anti-DNA Polymerase Kappa/POLK (4F12)

YF-MA18501 100 ug
EUR 363
Description: Mouse monoclonal to DNA Polymerase Kappa/POLK

DNA polymerase ? (Rat)

10-101 20ug
EUR 388
Description: The DNA polymerase ? (Rat) is available in Europe and for worldwide shipping via Gentaur.

DNA polymerase ? (Rat)

10-102 100ug
EUR 923
Description: The DNA polymerase ? (Rat) is available in Europe and for worldwide shipping via Gentaur.

Taq DNA Polymerase

9001-2500
EUR 512

Taq DNA Polymerase

9001-500
EUR 191

PFU DNA Polymerase

9003-2500
EUR 512

PFU DNA Polymerase

9003-500
EUR 180

Laq? DNA Polymerase

9004-2500
EUR 610

Laq? DNA Polymerase

9004-500
EUR 218

Taq DNA polymerase

BT10101 500U
EUR 80
Description: High quality Taq polymerase for different PCR variations and downstream applications.

Taq DNA polymerase

BT10102 1000U
EUR 109
Description: High quality Taq polymerase for different PCR variations and downstream applications.

Taq DNA polymerase

BT10103 2000U
EUR 167
Description: High quality Taq polymerase for different PCR variations and downstream applications.

HotTaq DNA polymerase

BT10201 500U
EUR 120
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.

HotTaq DNA polymerase

BT10202 1000U
EUR 190
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.

Taq DNA Polymerase

BA00103 500U
EUR 87
Description: High quality Taq polymerase for different PCR variations and downstream applications.

Taq DNA Polymerase

BA00104 1000U
EUR 123
Description: High quality Taq polymerase for different PCR variations and downstream applications.

Taq DNA Polymerase

BA00105 2500U
EUR 232
Description: High quality Taq polymerase for different PCR variations and downstream applications.

HotTaq DNA Polymerase

BA00203 500U
EUR 145
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.

HotTaq DNA Polymerase

BA00204 1000U
EUR 239
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.

HotTaq DNA Polymerase

BA00205 2500U
EUR 522
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.

RedTaq DNA Polymerase

BA00303 500U
EUR 123
Description: High quality RedTaq polymerase for different PCR variations and downstream applications.

RedTaq DNA Polymerase

BA00304 1000U
EUR 196
Description: High quality RedTaq polymerase for different PCR variations and downstream applications.

RedTaq DNA Polymerase

BA00305 2500U
EUR 413
Description: High quality RedTaq polymerase for different PCR variations and downstream applications.

Pfu DNA Polymerase

BA00503 500U
EUR 145
Description: High quality Pfu polymerase for different PCR variations and downstream applications.

Pfu DNA Polymerase

BA00504 1000U
EUR 239
Description: High quality Pfu polymerase for different PCR variations and downstream applications.

Pfu DNA Polymerase

BA00505 2500U
EUR 522
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

Kodaq DNA Polymerase

G498 250 U (50 ul)
EUR 87

Kodaq DNA Polymerase

G499 1000 U (200 ul)
EUR 158

AceTaq DNA Polymerase

P401-d1 250 U
EUR 136

AceTaq DNA Polymerase

P401-d2 1000 U
EUR 227

AceTaq DNA Polymerase

P401-d3 3000 U
EUR 447

Taq DNA Polymerase

L7051001 1000 unit
EUR 195
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 87
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.

Ready? DNA Polymerase

M1146-1000
EUR 332

Ready? DNA Polymerase

M1146-10000
EUR 1066

Ready? DNA Polymerase

M1146-5000
EUR 800

Breeze? DNA Polymerase

M1148-1000
EUR 1039

The relative abundance of actinobacteria was significantly lowered when using the Mobio Kit amplification method or fluidigm, and the relative abundance of firmoles was lower when using the Qiagen Kit. Microbial community profiles based on fluidigm-generated amplicon libraries have not been comparable to those generated by more commonly used methods. Researchers should carefully consider limitations and biases that different methods of extraction and amplification can introduce their results. In addition, a deeper benchmarking of automated methods and multiplexing is required to determine the magnitude of the potential compromise between the quality and the amount of data.

Leave A Comment