Tissue expression of rat ciliary neurotrophic factor (CNTF) mRNA and production of the recombinant CNTF.
We used the polymerase chain reaction to show that CNTF mRNA is widely expressed in the brain, heart, lung, liver, kidney and testis of the rat, in addition to preferential expression in the sciatic nerve. We produced the recombinant CNTF in E. coli using rat sciatic nerve cDNA and obtained high yields of pure CNTF. The biological activity of the recombinant protein was comparable to that of native rat CNTF in its support of the survival of cultured embryonic day-8 chick ciliary ganglions. Western Blot analysis using antibody against the recombinant CNTF clearly showed the presence of CNTF protein (25kD) in rabbit sciatic nerve.
Ohta
K
K
Clinical Research Center, Utano National Hospital, Kyoto, Japan.
Hara
H
H
Hayashi
K
K
Itoh
N
N
Ohi
T
T
Ohta
M
M
D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002642:Chick Embryo; D020934:Ciliary Neurotrophic Factor; D003001:Cloning, Molecular; D017931:DNA Primers; D004926:Escherichia coli; D005726:Ganglia, Parasympathetic / Q000166:cytology; D015870:Gene Expression; D007668:Kidney / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D009206:Myocardium / Q000378:metabolism; D009414:Nerve Growth Factors / Q000096:biosynthesis / Q000494:pharmacology; D009419:Nerve Tissue Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology; D009474:Neurons / Q000166:cytology / Q000187:drug effects; D009928:Organ Specificity; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology; D012584:Sciatic Nerve / Q000378:metabolism; D013737:Testis / Q000378:metabolism
D020934:Ciliary Neurotrophic Factor; D017931:DNA Primers; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins