Log In Sign up
The largest database of trusted experimental protocols

Pgbkt7 bait

Manufactured by Takara Bio
Visit Supplier

PGBKT7 is a plasmid vector used for yeast two-hybrid screening. It contains a GAL4 DNA-binding domain for generating bait constructs.

Automatically generated - may contain errors

Lab products found in correlation

Pcdna3.1d v5 his topo vector, by Thermo Fisher Scientific (1 mentions) Pgadt7 ad prey, by Takara Bio (1 mentions) Quikchange 2 xl, by Agilent Technologies (1 mentions)

Spelling variants of this product

PGBKT7 (413 citations) PGBKT7 bait vector (8 citations)

3 protocols using pgbkt7 bait

1

Yeast Two-Hybrid Screening of HIV-1 Gag

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
The infectious molecular clone pNL-CH [33 (link)], which was derived from the pNL4−3 clone of HIV-1, was a gift from R. Swanstrom. To perform Y2H screening, the Pr55gag and CA coding region of the HIV-1NL–CH were cloned into pGBKT7 (bait) (Clontech Laboratories, Inc.) as previously described [34 (link)].To prepare the ATE1-V5 expression vector, the coding region of ATE1, which was amplified by PCR using the primers ATE1-TCF (5′-AAAGCTTGCCATGGCTTTCTGGGCGGGG-3′) and ATE1-TCR (5′-TCTCGAGCAGTTTCTGAACAGCAGCATCCG-3′), was cloned into the pcDNA™ 3.1D/V5-His-TOPO® vector (Thermo Fisher Scientific, Inc.).
Kishimoto N., Okano R., Akita A., Miura S., Irie A., Takamune N, & Misumi S. (2021). Arginyl-tRNA-protein transferase 1 contributes to governing optimal stability of the human immunodeficiency virus type 1 core. Retrovirology, 18, 30.
+ Open protocol
+ Expand
2

Yeast Two-Hybrid Analysis of TEX11

Check if the same lab product or an alternative is used in the 5 most similar protocols
Human TEX11 truncations (DNA encoding amino acids 1–189, 1–417, 1–457, and 1–814) were cloned into pGADT7-AD (Prey, Clontech), to produce fusions to the Gal4 DNA-binding and activation domains. A plasmid containing full-length human SHOC1 was constructed by cloning the appropriate PCR product into pGBKT7 (Bait, Clontech). All fusions were confirmed by sequencing. Two-hybrid assays were conducted in the AH109 strain background. After mating, colonies containing both plasmids were selected using media lacking tryptophan and leucine. Interactions between partners were assayed by growth on synthetic media lacking tryptophan, leucine, adenine and histidine. Transformations were carried out according to the matchmaker kit manual (BD Biosciences).
Guiraldelli M.F., Felberg A., Almeida L.P., Parikh A., de Castro R.O, & Pezza R.J. (2018). SHOC1 is a ERCC4-(HhH)2-like protein, integral to the formation of crossover recombination intermediates during mammalian meiosis. PLoS Genetics, 14(5), e1007381.
+ Open protocol
+ Expand
3

Yeast Two-Hybrid Analysis of MLH1, MSH2, PMS2, and MSH6

Check if the same lab product or an alternative is used in the 5 most similar protocols
The introduction of variants in MLH1 (vector pCite-MLH1) and MSH2(vector pAL112) (primer sequences are available upon request) was performed according to the manufacturer’s protocol
(QuikChange II XL, Agilent Technologies). Substitutions were verified by sequencing.
Plasmid pCite-MLH1 (wild type or mutant) was digested with NdeI and
EcoRI and inserted into yeast two-hybrid (Y2H) vector pGBKT7 (bait, Clontech). MSH2 variants
were inserted into the NcoI site in pGK240. The coding sequences were subcloned into two-hybrid vector pGBKT7.
PMS2 was inserted in NdeI and SmaI sites of vector pGK239 (pGADT7 backbone,
prey). MSH6 was subcloned from pGEX-MSH6 into pGADT7.
Y2H analysis was set up according to the manufacturer’s protocol (Clontech). The Y2HGold yeast strain was
cotrans-formed with PMS2/MLH1- or MSH2/MSH6-expressing plasmids. Plates were grown for 3–5 days. The PMS2wild type, MLH1 wild type, and MLH1 mutants were plated on plates lacking Trp/Leu and containing
X-α-gal, and plates lacking Trp/Leu/His/Ada and containing X-α-gal and Aureobasidin A. The MSH6wild type, MSH2 wild type, and MSH2 mutants were plated on plates lacking Trp/Leu and containing
X-α-gal and on plates lacking Trp/Leu/His and containing X-α-gal and Aureobasidin A. Colonies were counted and
scored for growth and for blue (interaction)/white (no interaction).
Drost M., Tiersma Y., Thompson B.A., Frederiksen J.H., Keijzers G., Glubb D., Kathe S., Osinga J., Westers H., Pappas L., Boucher K.M., Molenkamp S., Zonneveld J.B., van Asperen C.J., Goldgar D.E., Wallace S.S., Sijmons R.H., Spurdle A.B., Rasmussen L.J., Greenblatt M.S., de Wind N, & Tavtigian S.V. (2018). A functional assay-based procedure to classify mismatch repair gene variants in Lynch syndrome. Genetics in medicine : official journal of the American College of Medical Genetics, 21(7), 1486-1496.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!