Identification of chicken calbindin D28K pre-messenger RNA sequences by polymerase chain reaction.
A transcribed RNA sequence encompassing the junction between the first intron and the second exon of the chicken calbindin D28K gene was copied in a cDNA fragment and subsequently amplified by polymerase chain reaction. When intestinal RNA is used as template, the appearance of the 161 bp amplified fragment is strictly dependent on the vitamin D status of the animal. In fact no amplified fragment is obtained when the RNA is extracted from the intestine of vitamin D-deficient chickens, while it is easily detected when the RNA is extracted only 30 min after injection with 1,25-dihydroxycholecalciferol. Conversely, the amplified fragment is obtained, irrespectively of the vitamin D status of the animal, when the RNA template is extracted from the brain. The appearance of unspliced RNA sequences upon vitamin D induction is followed, after a 30 min lag, by the appearance of the corresponding mature mRNA sequences.
Ferrari
S
S
Istituto di Chimica Biologica, Università di Modena, Italy.
Battini
R
R
D000818:Animals; D001483:Base Sequence; D001923:Brain Chemistry; D064026:Calbindins; D002645:Chickens; D007422:Intestines / Q000032:analysis; D008969:Molecular Sequence Data; D009694:Nucleic Acid Precursors / Q000032:analysis*; D016133:Polymerase Chain Reaction; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000032:analysis*; D064030:S100 Calcium Binding Protein G / Q000235:genetics*; D014158:Transcription, Genetic; D014808:Vitamin D Deficiency / Q000235:genetics
D064026:Calbindins; D009694:Nucleic Acid Precursors; D012333:RNA, Messenger; D064030:S100 Calcium Binding Protein G