Cloning and expression of rat GFAP cDNA in Escherichia coli.
A cDNA fragment encoding GFAP was amplified by reverse transcription PCR from total mRNA isolated from primary culture of rat astrocytes and cloned for expression in Escherichia coli using pET-28a vector. High level of GFAP expression was confirmed by SDS-PAGE, while immunochemical identity was verified by immunoblotting. The constructed producer strain is a cheap source of GFAP and can be used for diagnostic purposes.
Chekhonin
V P
VP
Laboratory of Immunochemistry, Department of Biological Psychiatry, V. P. Serbskii Center of Social and Forensic Psychiatry, Moscow.
Pavlov
K A
KA
Shkoporov
A N
AN
Yefimov
B A
BA
Gurina
O I
OI
Dmitrieva
T B
TB
D000818:Animals; D001253:Astrocytes / Q000378:metabolism; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis*; D004926:Escherichia coli / Q000378:metabolism*; D005904:Glial Fibrillary Acidic Protein / Q000096:biosynthesis* / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis
D018076:DNA, Complementary; D005904:Glial Fibrillary Acidic Protein; D012333:RNA, Messenger; D011994:Recombinant Proteins