Author/Editor		Debeljak, Nataša
Title		Mouse as a model organism to study the role of the Cyp51 gene in spermatogenesis
Translated title		Miš kot modelni organizem za študij vloge gena CYP51 v spermatogenezi
Type		monografija
Place		Ljubljana
Publisher		Faculty of medicine
Publication year		2002
Volume		str. 130
Language		eng
Abstract		A new member of the CYP51 family, murine lanosterol 14a-demethylase (Cyp51), has been characterised. The mouse Cyp51 gene is aproximately 23-kb long with 10 exons and is encoding a 503 amino acids long protein. Similar gene organisation, conserved promoter region and over 91 % amino acid identity show a high similarity to other mammalian CYP51 genes. Comparison of all known CYP51 proteins by the neighbour-joining method suggests that fungal and animal CYP51 genes arose from a common ancestral gene and interestingly, that plant and bacterial CYP51 genes share a common progenitor. A single copy of the mouse Cyp51 gene resides on chromosome 5, region A2, close to the centromere. No signals outside this Cyp51 region were detected as well as no evidence of Cyp51 processed pseudogenes was found. Further the development of a Cyp51 male germ-cell specific knockout mouse took place to address the in vivo role of the gene in mammalian reproduction. A total knockout would most probably be lethal as CYP51 is involved in cholesterol biosynthesis, thus Cre-IoxP strategy was used for preparation of a conditional knockout. Cyp51loxP recombinant targeting vectors containing two isolated IoxP sites in introns 2 and 4 and IoxP- or Frt-flanked selection cassette were prepared by subcloning murine Cyp51 regions. The IoxP-flanked Cyp51 fragment (containing introns 2-4) will be deleted after expressing Cre recombinase, which should lead to a stžift of the reading frame in the translated CYP51 protein and to introduction of a stop codon after amino acid 101, resulting in the nonfunctional CYP51. (Abstract truncated at 2000 charactes).
Descriptors		TESTIS SPERMATOGENESIS CYTOCHROME P-450 LANOSTEROL RECOMBINATION, GENETIC POLYMERASE CHAIN REACTION MICE, TRANSGENIC PROMOTER REGIONS (GENETICS) GERM CELLS IN SITU HYBRIDIZATION, FLUORESCENCE CLONING, MOLECULAR BASE SEQUENCE