cDNA Cloning and Functional Characterization of Rat Intestinal Monocarboxylate Transporter

1995 ◽  
Vol 217 (1) ◽  
pp. 370-377 ◽  
Author(s):  
H. Takanaga ◽  
I. Tamai ◽  
S. Inaba ◽  
Y. Sai ◽  
H. Higashida ◽  
...  
Keyword(s):  
Cdna Cloning ◽  
Functional Characterization ◽  
Monocarboxylate Transporter

cDNA cloning, expression, and functional characterization of a zebrafish SULT1 cytosolic sulfotransferase

2003 ◽  
Vol 414 (1) ◽  
pp. 67-73 ◽  
Author(s):  
Takuya Sugahara ◽  
Chau-Ching Liu ◽  
Glendora Carter ◽  
T. Govind Pai ◽  
Ming-Cheh Liu
Keyword(s):  
Cdna Cloning ◽  
Functional Characterization

Design and Characterization of a Fluorescent Reporter Enabling Live-cell Monitoring of MCT8 Expression

2021 ◽  
Author(s):  
Adina Sophie Graffunder ◽  
Sarah Paisdzior ◽  
Robert Opitz ◽  
Kostja Renko ◽  
Peter Kühnen ◽  
...  
Keyword(s):  
Expression Patterns ◽  
Functional Characterization ◽  
Live Cell ◽  
Monocarboxylate Transporter ◽  
Start Codon ◽  
Future Application ◽  
Gene Assay ◽  
Cell Monitoring

AbstractThe monocarboxylate transporter 8 (MCT8) is a specific thyroid hormone transporter and plays an essential role in fetal development. Inactivating mutations in the MCT8 encoding gene SLC16A2 (solute carrier family 16, member 2) lead to the Allan-Herndon-Dudley syndrome, a condition presenting with severe endocrinological and neurological phenotypes. However, the cellular distribution pattern and dynamic expression profile are still not well known for early human neural development. Objective Development and characterization of fluorescent MCT8 reporters that would permit live-cell monitoring of MCT8 protein expression in vitro in human induced pluripotent stem cell (hiPSC)-derived cell culture models. Methods A tetracysteine (TC) motif was introduced into the human MCT8 sequence at four different positions as binding sites for fluorescent biarsenical dyes. Human Embryonic Kidney 293 cells were transfected and stained with fluorescein-arsenical hairpin-binder (FlAsH). Counterstaining with specific MCT8 antibody was performed. Triiodothyronine (T3) uptake was indirectly measured with a T3 responsive luciferase-based reporter gene assay in Madin-Darby Canine Kidney 1 cells for functional characterization. Results FlAsH staining and antibody counterstaining of all four constructs showed cell membrane expression of all MCT8 constructs. The construct with the tag after the first start codon demonstrated comparable T3 uptake to the MCT8 wildtype. Conclusion Our data indicate that introduction of a TC-tag directly after the first start codon generates a MCT8 reporter with suitable characteristics for live-cell monitoring of MCT8 expression. One promising future application will be generation of stable hiPSC MCT8 reporter lines to characterize MCT8 expression patterns during in vitro neuronal development.

scholarly journals cDNA Cloning, Expression, and Functional Characterization of PI31, a Proline-rich Inhibitor of the Proteasome

2000 ◽  
Vol 275 (24) ◽  
pp. 18557-18565 ◽  
Author(s):  
Sandra L. McCutchen-Maloney ◽  
Koichi Matsuda ◽  
Naoki Shimbara ◽  
Derk D. Binns ◽  
Keiji Tanaka ◽  
...  
Keyword(s):  
Cdna Cloning ◽  
Functional Characterization

scholarly journals cDNA cloning and expression of rat homeobox gene, Hex, and functional characterization of the protein

1999 ◽  
Vol 339 (1) ◽  
pp. 111 ◽  
Author(s):  
Takashi TANAKA ◽  
Tetsuya INAZU ◽  
Kazuya YAMADA ◽  
Zaw MYINT ◽  
Vincent W. KENG ◽  
...  
Keyword(s):  
Cdna Cloning ◽  
Functional Characterization ◽  
Homeobox Gene ◽  
Cloning And Expression
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