Author/Editor		Grubič, Zoran; Komel, Rado
Title		Synthesis of the DNA probe for the determination of rat AChE mRNA
Type		članek
Source		Chem Biol Interact
Vol. and No.		Letnik 87, št. 1-3
Publication year		1993
Volume		str. 245-8
Language		eng
Abstract		This is a preliminary report on our attempts of synthesis by polymerase chain reaction (PCR), the cDNA probe for the determination of mRNA of the AChE catalytic subunit. As our strategy we took the advantage of the fact that sequence identity of AChE gene increases with phylogenetic proximity. Single codon usage could therefore be applied. Two non-degenerate PCR primers were synthesised corresponding to AChE regions which were highly conservative among species analyzed until now. The sequence amplified by these two primers should be 339 base pairs long as concluded from mouse AChE sequence. By determining the nucleotide sequence of the PCR product and by comparison of this sequence with the corresponding mouse AChE region, we would be able to verify the correspondence of our PCR product to the rat AChE gene fragment. Only the first four amino acids of our PCR product flanking Phe 200, which is the first amino acid from the A2 primer, are 100 percent homologous with the mouse AChE. However, from the next 18 amino acids towards the N-terminal, only 4 are homologous with the mouse AChE. Since we expected more than 90 percent homology between the phylogenetically closely related species of mouse and rat, we doubt that the DNA sequence obtained belongs to the rat AChE gene.
Descriptors		ACETYLCHOLINESTERASE DNA PROBES RNA, MESSENGER AMINO ACID SEQUENCE BASE SEQUENCE GENE AMPLIFICATION MICE MOLECULAR SEQUENCE DATA POLYMERASE CHAIN REACTION RATS RNA, MESSENGER SEQUENCE HOMOLOGY, AMINO ACID