Exp Mol Med.  2000 Sep;32(3):115-119.

Molecular cloning and sequencing of rat Cdc42 GTPase cDNA

• Han JS
• Kim JH
• Kim JG
• Park JB
• Noh DY
• Lee KH

Abstract

Cdc42 is a member of the Rho family of small GTP-ase and plays an important role in intracellular signaling pathways regulating cell morphology, motility and stimulation of DNA synthesis. We have isolated cDNA encoding Cdc42 from a rat brain cDNA library using PCR-cloning strategy. The sequence of isolated gene revealed an open reading frame of 576 nucleotides with the potential to encode a protein of 191 amino acids with a predicted molecular weight of 21 kD. The resulting sequence was incorporated into the GenBank with accession number, AF205635. Sequence analysis revealed that overall cDNA sequence identity is 96% with human G25K and 52% with rat Chp, a homologue of the GTPase human Cdc42Hs, and having one nucleotide difference from the mouse Cdc42. However, putative protein sequence was identical to the mouse and human brain Cdc42Hs. On expression of the cDNA in COS-7 cells, a protein molecular weight of 21 kD was detected in immunoblotting using anti-human Cdc42 antibodies. Therefore, these results suggest that the cDNA we are reporting is most likely the rat homologue of the GTPase human Cdc42.

Keyword

GTPase; Cdc42; PCR Cloning; COS-7 cell; Rho

MeSH Terms

Amino Acid Sequence
Animal
Base Sequence
Cloning, Molecular
Comparative Study
Cross Reactions
DNA, Complementary/genetics
Human
Molecular Sequence Data
Rats
Sequence Analysis, DNA
Sequence Homology, Amino Acid
cdc42 GTP-Binding Protein/immunology
cdc42 GTP-Binding Protein/genetics*