[Expression of a gene fragment from the alpha-subunit of the acetylcholine receptor from Torpedo californica in Saccharomyces cerevisiae].
A fragment of the AChR alpha-subunit gene from Torpedo californica (about 800 bp) was joined in frame with a synthetic duplex, coding for a changed leader peptide of the Kluyveromyces lactis killer toxin under the control of the reconstructed glyceraldehyde-3-phosphate dehydrogenase gene promoter in the pUC8 vector. Translation termination codons in all frames were inserted as a part of a self-complementary oligodeoxynucleotide to the Eco47III site (807 bp from the start of the gene). Vector pJDAch was constructed by cloning the expression cassette between the BamHI and HindIII sites in the multicopy yeast plasmid pJDB207. Yeast cells harbouring the pJDAch vector produced mainly an N-glycosylated fragment of the acetylcholine receptor alpha-subunit, detected by means of [125I]-alpha-bungarotoxin. The fragment's location in the membrane fraction after disruption of the yeast cells simplified its isolation.
Teliakova
O V
OV
Utkin
Iu N
IuN
Tsetlin
V I
VI
Severtsova
I V
IV
Zvonok
N M
NM
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D005822:Genetic Vectors; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D055646:Killer Factors, Yeast; D008969:Molecular Sequence Data; D009183:Mycotoxins / Q000235:genetics*; D009838:Oligodeoxyribonucleotides; D010446:Peptide Fragments / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011950:Receptors, Cholinergic / Q000737:chemistry / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013728:Terminator Regions, Genetic; D014101:Torpedo / Q000378:metabolism*
D055646:Killer Factors, Yeast; D009183:Mycotoxins; D009838:Oligodeoxyribonucleotides; D010446:Peptide Fragments; D011950:Receptors, Cholinergic; C058975:zymocin; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases
Ekspressiia fragmenta gena alpha-sub''edinitsy atsetilkholinovogo retseptora iz Torpedo californica v Saccharomyces cerevisiae.