[Cloning, genomic organization and promoter activity of the mouse zinc finger protein gene ZF-12].
The human zinc finger protein ZNF191 is a krüppel-like transcription factor, which may be relevant to many diseases such as neuropsychiatric, cardiovascular and liver caner diseases. To elucidate the function of ZNF191 by gene targeting, it is necessary to clone and characterize of the homologous gene in model organisms (mice). The mouse homologous gene (ZF-12) was cloned and sequenced for the first time, the GenBank accession number is AY052495. It contains four exons and three introns; all intronic splice sites exhibited consensus GT/AG sequences. The single nucleotide polymorphisms (SNPs) in exon 2 and the alternative length of 3'-untranslated region (3'-UTR) have been found. The linkage of the ZF-12 gene and the zinc finger protein gene Zfp-35 has been found, so the ZF-12 gene can be localized to B3 to C or beside of chromosome 18. We assessed approximately 1.2 kb of 5'-flanking region of the ZF-12 gene for basal promoter activity. A series of deletion mutants of 5'-flanking region linked to the luciferase gene was constructed. Basal level expression of these constructs was tested in COS-7 cells, NIH3T3 cells and HeLa cells. By measuring luciferase activity, which was transiently expressed in the transfected cells, we found that regulatory elements sufficient for basal expression lie between -762 and +70 bp relative to the transcription start site and that a negative regulatory region lie between -824 and -762 bp. This research provides a basis for further study on ZF-12 by gene targeting.
Li
Jian-Zhong
JZ
Department of Medical Genetics, Second Military University, Shanghai 200433, China.
Chen
Xia
X
Wang
Shui-Liang
SL
Sun
Xia
X
Zhang
Ya-Zhou
YZ
Yu
Long
L
Fu
Ji-Liang
JL
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002352:Carrier Proteins / Q000235:genetics*; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry / Q000235:genetics; D005091:Exons; D015870:Gene Expression; D005796:Genes / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D007438:Introns; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA
D002352:Carrier Proteins; D011993:Recombinant Fusion Proteins; C474810:Zfp24 protein, mouse; D004247:DNA; D008156:Luciferases