Molecular cloning and characterization of a ras p21-like GTP-binding protein (24KG) from rat liver.
We have isolated cDNA clones from a rat liver cDNA library that encode a ras p21-like small GTP-binding protein (24KG) which was purified from the microsomes-Golgi complex fraction of the rat liver. The cloning was accomplished using polymerase chain reaction amplified with a set of oligonucleotide primers which were designed from the partial amino acid sequences for 24KG. The cDNA contained an open reading frame encoding a 216 amino acid protein with a calculated Mr weight of 24,397. This Mr weight was similar to that of the purified 24KG estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The sequence analysis of 24KG revealed that a 24KG cDNA is the rat counterpart of a rab11 cDNA cloned from a Madin-Darby canine kidney cell cDNA library. The 1.0-kilobase 24KG mRNA corresponding to the isolated cDNA was also detected in various rat tissues, such as brain, testis, spleen, and heart.
Sakurada
K
K
Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., Japan.
Uchida
K
K
Yamaguchi
K
K
Aisaka
K
K
Ito
S
S
Ohmori
T
T
Takeyama
Y
Y
Ueda
T
T
Hori
Y
Y
Ohyanagi
H
H
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D015342:DNA Probes; D019204:GTP-Binding Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015723:Gene Library; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D011061:Poly A / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics*; D012313:RNA / Q000235:genetics; D012333:RNA, Messenger; D051381:Rats; D011919:Rats, Inbred Strains; D012689:Sequence Homology, Nucleic Acid
D015342:DNA Probes; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D011061:Poly A; D012313:RNA; D004247:DNA; D019204:GTP-Binding Proteins; D016283:Proto-Oncogene Proteins p21(ras)