Uses of lacZ to Study Gene Function: Evaluation of β-Galactosidase Assays Employed in the Yeast Two-Hybrid System

2000 ◽  
Vol 285 (1) ◽  
pp. 1-15 ◽  
Author(s):  
Ilya G. Serebriiskii ◽  
Erica A. Golemis
Keyword(s):  
Hybrid System ◽  
Gene Function ◽  
Function Evaluation ◽  
Yeast Two Hybrid ◽  
Yeast Two Hybrid System ◽  
Two Hybrid

Electrophoretic Cell Manipulation and Electrochemical Gene-Function Analysis Based on a Yeast Two-Hybrid System in a Microfluidic Device

2008 ◽  
Vol 80 (10) ◽  
pp. 3722-3727 ◽  
Author(s):  
Tomoyuki Yasukawa ◽  
Kuniaki Nagamine ◽  
Yoshiko Horiguchi ◽  
Hitoshi Shiku ◽  
Masahiro Koide ◽  
...  
Keyword(s):  
Microfluidic Device ◽  
Hybrid System ◽  
Gene Function ◽  
Function Analysis ◽  
Cell Manipulation ◽  
Yeast Two Hybrid ◽  
Yeast Two Hybrid System ◽  
Gene Function Analysis ◽  
Two Hybrid

Expression and Construction of Yeast Expression Vector Containing CsTCTP1 Gene from Cucumber in Yeast Two-Hybrid System

2017 ◽  
Vol 10 (1) ◽  
pp. 55-61
Author(s):  
Qiu-Min Chen ◽  
Na Cui ◽  
Yang Yu ◽  
Xiang-Nan Meng ◽  
Hai-Yan Fan
Keyword(s):  
Hybrid System ◽  
Gene Function ◽  
Expression Vector ◽  
Detection System ◽  
Yeast Expression ◽  
Yeast Two Hybrid ◽  
Bait Protein ◽  
Yeast Two Hybrid System ◽  
Hybrid Detection ◽  
Two Hybrid

Background: The translationally controlled tumor protein (TCTP) was originally found in tumor tissue, and later found in other tissues. Initially, TCTP was considered a kind of growth-associated protein. Recent studies have shown that TCTP has many biological functions. Objective: To verification of CsTCTP1 gene function by yeast two-hybrid system, the pGBKT7- CsTCTP1 yeast expression vector was constructed and cytotoxicity and self-activating activity were detected, which could lay the foundation for further studies on gene function and make a preparation for verification of CsTCTP1 gene function by yeast two-hybrid system. Method: Specific PCR, conventional sequencing, heat shock conversion method and TE/LiAC transformation method. Results: We constructed a yeast expression vector containing the CsTCTP1 gene. The CsTCTP1 coding sequence was inserted into a pGBKT7 vector as a bait protein and then transformed into the Y2HGold yeast stain. Conclusion: We found that CsTCTP1 protein had no cytotoxic effect and could not be self-activated. The constructed bait expression vector can be used in the subsequent yeast two - hybrid detection system.

Identifications of DNA Sequences Encoding Key Region of SCR Interacting with SRK Extracellular Domain by Using Yeast Two-Hybrid System

2013 ◽  
Vol 38 (9) ◽  
pp. 1583-1591
Author(s):  
Li-Yan XUE ◽  
Bing LUO ◽  
Li-Quan ZHU ◽  
Yong-Jun YANG ◽  
He-Cui ZHANG ◽  
...  
Keyword(s):  
Hybrid System ◽  
Dna Sequences ◽  
Extracellular Domain ◽  
Yeast Two Hybrid ◽  
Yeast Two Hybrid System ◽  
Two Hybrid

Screening of binding proteins that interact with human Salvador 1 in a human fetal liver cDNA library by the yeast two-hybrid system

2012 ◽  
Vol 39 (8) ◽  
pp. 8225-8230 ◽  
Author(s):  
Xiaolan Li ◽  
Xuelai Luo ◽  
Zhaoming Li ◽  
Guihua Wang ◽  
Hui Xiao ◽  
...  
Keyword(s):  
Cdna Library ◽  
Hybrid System ◽  
Binding Proteins ◽  
Fetal Liver ◽  
Yeast Two Hybrid ◽  
Human Fetal Liver ◽  
Yeast Two Hybrid System ◽  
Liver Cdna Library ◽  
Two Hybrid

Identification of MAL2, a Novel Member of the MAL Proteolipid Family, Though Interactions with TPD52-like Proteins in the Yeast Two-Hybrid System

Genomics ◽  
2001 ◽  
Vol 76 (1-3) ◽  
pp. 81-88 ◽  
Author(s):  
Sarah H.D Wilson ◽  
Angela M Bailey ◽  
Craig R Nourse ◽  
Marie-Geneviève Mattei ◽  
Jennifer A Byrne
Keyword(s):  
Hybrid System ◽  
Yeast Two Hybrid ◽  
Yeast Two Hybrid System ◽  
Two Hybrid

scholarly journals Interactions among members of the Bcl-2 protein family analyzed with a yeast two-hybrid system.

1994 ◽  
Vol 91 (20) ◽  
pp. 9238-9242 ◽  
Author(s):  
T. Sato ◽  
M. Hanada ◽  
S. Bodrug ◽  
S. Irie ◽  
N. Iwama ◽  
...  
Keyword(s):  
Hybrid System ◽  
Protein Family ◽  
Yeast Two Hybrid ◽  
Yeast Two Hybrid System ◽  
Two Hybrid

scholarly journals Subunit Interactions in the mariner Transposase

Genetics ◽  
1996 ◽  
Vol 144 (3) ◽  
pp. 1087-1095 ◽  
Author(s):  
Allan R Lohe ◽  
David T Sullivan ◽  
Daniel L Hartl
Keyword(s):  
Hybrid System ◽  
Catalytic Domain ◽  
Wild Type ◽  
Target Element ◽  
Yeast Two Hybrid ◽  
Subunit Interactions ◽  
Hsp70 Promoter ◽  
Yeast Two Hybrid System ◽  
Transposition Reaction ◽  
Two Hybrid

Abstract We have studied the Mos1 transposase encoded by the transposable element mariner. This transposase is a member of the “D,D(35)E” superfamily of proteins exhibiting the motif D,D(34)D. It is not known whether this transposase, or other eukaryote transposases manifesting the D,D(35)E domain, functions in a multimeric form. Evidence for oligomerization was found in the negative complementation of Mos1 by an EMS-induced transposase mutation in the catalytic domain. The transposase produced by this mutation has a glycine-to-arginine replacement at position 292. The G292R mutation strongly interferes with the ability of wild-type transposase to catalyze excision of a target element. Negative complementation was also observed for two other EMS mutations, although the effect was weaker than observed with G292R. Results from the yeast two-hybrid system also imply that Mos1 subunits interact, suggesting the possibility of subunit oligomerization in the transposition reaction. Overproduction of Mos1 subunits through an hsp70 promoter also inhibits excision of the target element, possibly through autoregulatory feedback on transcription or through formation of inactive or less active oligomers. The effects of both negative complementation and overproduction may contribute to the regulation of mariner transposition.

ralGDS family members interact with the effector loop of ras p21

1994 ◽  
Vol 14 (11) ◽  
pp. 7483-7491
Author(s):  
A Kikuchi ◽  
S D Demo ◽  
Z H Ye ◽  
Y W Chen ◽  
L T Williams
Keyword(s):  
Hybrid System ◽  
Family Members ◽  
Dominant Negative Mutant ◽  
Yeast Two Hybrid ◽  
Amino Acid Homology ◽  
Yeast Two Hybrid System ◽  
Ras P21 ◽  
Two Hybrid ◽  
Interacting Domain

Using a yeast two-hybrid system, we identified a novel protein which interacts with ras p21. This protein shares 69% amino acid homology with ral guanine nucleotide dissociation stimulator (ralGDS), a GDP/GTP exchange protein for ral p24. We designated this protein RGL, for ralGDS-like. Using the yeast two-hybrid system, we found that an effector loop mutant of ras p21 was defective in interacting with the ras p21-interacting domain of RGL, suggesting that this domain binds to ras p21 through the effector loop of ras p21. Since ralGDS contained a region highly homologous with the ras p21-interacting domain of RGL, we examined whether ralGDS could interact with ras p21. In the yeast two-hybrid system, ralGDS failed to interact with an effector loop mutant of ras p21. In insect cells, ralGDS made a complex with v-ras p21 but not with a dominant negative mutant of ras p21. ralGDS interacted with the GTP-bound form of ras p21 but not with the GDP-bound form in vitro. ralGDS inhibited both the GTPase-activating activity of the neurofibromatosis gene product (NF1) for ras p21 and the interaction of Raf with ras p21 in vitro. These results demonstrate that ralGDS specifically interacts with the active form of ras p21 and that ralGDS can compete with NF1 and Raf for binding to the effector loop of ras p21. Therefore, ralGDS family members may be effector proteins of ras p21 or may inhibit interactions between ras p21 and its effectors.

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