Chorionic villus cDNA library displays expression of butyrylcholinesterase: putative genetic disposition for ecological danger.
Gene expression in chorionic villi may be particularly vulnerable to environmental exposure to poisonous substances. To reveal villus gene products which are thus subject to poisoning, molecular cloning was employed. A single sample of apparently normal chorionic villi (approximately 40 mg, from 9 weeks' gestation) was microscopically dissected to ensure purity of fetal tissue. Total RNA was extracted by isothiocyanate and directly employed for reverse transcription. A chorionic villus cDNA library was constructed from this preparation in the phage vector lambda gt10 and contained 60,000 independent recombinants. In the present study, this cDNA library was screened with labelled cDNA probes encoding human butyrylcholinesterase (BCHE) and acetylcholinesterase (ACHE). Nine BCHEcDNA clones were isolated out of 1.6 x 10(6) phages (5.7 x 10(-6) of screened recombinants) and exhibited similar restriction patterns to those observed for BCHEcDNA from other human tissues. In contrast, no ACHEcDNA clones could be found in 4.0 x 10(6) screened phages (less than 2.5 x 10(-6) of recombinants). These findings demonstrate efficient transcription (similar to fetal brain levels) from the BCHE gene but not from the ACHE gene in chorionic villi, and support the notion that BCHE is involved in chorionic villus growth and development.
Zakut
H
H
Department of Obstetrics and Gynecology, Sackler Faculty of Medicine, Tel-Aviv University, Edith Wolfson Medical Center, Holon, Israel.
Lieman-Hurwitz
J
J
Zamir
R
R
Sindell
L
L
Ginzberg
D
D
Soreq
H
H
D000110:Acetylcholinesterase / Q000378:metabolism; D010582:Bacteriophage lambda; D002091:Butyrylcholinesterase / Q000096:biosynthesis* / Q000235:genetics; D002824:Chorionic Villi / Q000201:enzymology*; D003001:Cloning, Molecular; D015342:DNA Probes; D004587:Electrophoresis, Agar Gel; D004781:Environmental Exposure; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D015723:Gene Library; D006801:Humans; D009943:Organophosphorus Compounds / Q000494:pharmacology*; D012150:Polymorphism, Restriction Fragment Length; D012313:RNA / Q000302:isolation & purification; D015183:Restriction Mapping; D014158:Transcription, Genetic
D015342:DNA Probes; D009943:Organophosphorus Compounds; D012313:RNA; D000110:Acetylcholinesterase; D002091:Butyrylcholinesterase