TY - JOUR
T1 - Design of oligonucleotide probes for molecular cloning of β and γ subunits
AU - Gallagher, Christine
AU - Gautam, Narasimhan
N1 - Funding Information:
N.G. is an Established Investigator of the American Heart Association. This work was supported by grants from the National Institutes of Health and Monsanto-Searle.
PY - 1994/1/1
Y1 - 1994/1/1
N2 - Molecular cloning of cDNAs encoding distinct G-protein β- and γ- subunit types has shown that these subunits comprise families of proteins in mammalian tissues, cDNAs encoding four different β subunits and seven different γ subunits have now been identified. All the cDNAs were isolated using appropriate oligonucleotide probes. Two broadly different approaches have been used to isolate the cDNAs. The first method involves screening a cDNA library with oligonucleotide mixtures specific to the amino acid sequence of a protein. The second method is based on the polymerase chain reaction (PCR). In this method oligonucleotide mixtures specific to the amino acid sequence of a protein or specific to conserved amino acid residues among members of a family of proteins are used as PCR primers for amplifying a portion of the cDNA. Analysis with specific antibodies has revealed that purified G-protein βγ protein complexes from different sources are mixtures of β and γ subtypes. In addition, subtypes exist that are distinct from those for which cDNAs have been identified.
AB - Molecular cloning of cDNAs encoding distinct G-protein β- and γ- subunit types has shown that these subunits comprise families of proteins in mammalian tissues, cDNAs encoding four different β subunits and seven different γ subunits have now been identified. All the cDNAs were isolated using appropriate oligonucleotide probes. Two broadly different approaches have been used to isolate the cDNAs. The first method involves screening a cDNA library with oligonucleotide mixtures specific to the amino acid sequence of a protein. The second method is based on the polymerase chain reaction (PCR). In this method oligonucleotide mixtures specific to the amino acid sequence of a protein or specific to conserved amino acid residues among members of a family of proteins are used as PCR primers for amplifying a portion of the cDNA. Analysis with specific antibodies has revealed that purified G-protein βγ protein complexes from different sources are mixtures of β and γ subtypes. In addition, subtypes exist that are distinct from those for which cDNAs have been identified.
UR - http://www.scopus.com/inward/record.url?scp=0028092935&partnerID=8YFLogxK
U2 - 10.1016/S0076-6879(94)37084-2
DO - 10.1016/S0076-6879(94)37084-2
M3 - Article
C2 - 7935021
AN - SCOPUS:0028092935
SN - 0076-6879
VL - 237
SP - 471
EP - 482
JO - Methods in enzymology
JF - Methods in enzymology
IS - C
ER -