Generation of BPOZ gene knock-out mouse model

You Gui Xiang, Xia Sun, Long Wang, Lan Zhen Yan, Hua Yang, Wei Liu, Yong Xu, Guo Jiang Xu, Yi Wang, Jian Fei, Ji Liang Fu, Zhu Gang Wang

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

BPOZ is one protein containing ankyrin repeat and BTB (POZ) domain which gene is located at 3q21.3 in human genome. It was found that the expression of BPOZ gene is down-regulated in human ovary tumors, suggesting it is potentially a cell growth or tumor suppressor gene. BPOZ gene knockout mouse model was established for further in vivo study of its normal function and the role of BPOZ in tumorigenesis. The mouse genomic DNA sequence of BPOZ gene was verified through bioinformatic analysis. According to the BPOZ genomic DNA sequence, the strategy of gene targeting and constructing of knockout vector (XpPNT-BPOZ) were established to delete the region of mouse genome, which spans from exon 2 to 12 of BPOZ gene. The gene knockout vector XpPNT-BPOZ was constructed and confirmed by restriction enzyme digestion and sequencing. Electroporation of ES cells with XpPNT-BPOZ and screening of both G418 and Ganciclovoir resistant clones were performed according to common protocol. The homologous recombined ES cell clones were identified by PCR and confirmed by Southern blot analysis. After transplantation of homologous recombined ES cells into blastocysts through microinjection and mating of chimeras with C57BL/6J mice, 30 offspring with Aguoti fur in color were acquired. 15 of them (50%) show genotype heterozygous for BPOZ. The BPOZ heterozygotes were intercrossed to generate mutants homozygous. Finally, the cohort of mutants homozygous for BPOZ was established. The mutants are fertile, and further phenotype analysis is under way.

Original languageEnglish (US)
Pages (from-to)616-621
Number of pages6
JournalProgress in Biochemistry and Biophysics
Volume31
Issue number7
StatePublished - Jul 2004

Fingerprint

Gene Knockout Techniques
Knockout Mice
Genes
Tumor Suppressor Genes
Clone Cells
Ankyrin Repeat
Electroporation
Gene Targeting
Homologous Transplantation
Microinjections
Blastocyst
Human Genome
Heterozygote
Southern Blotting
Computational Biology
Inbred C57BL Mouse
Digestion
Ovary
Exons
Carcinogenesis

Keywords

  • BPOZ gene
  • Cell growth suppressor gene
  • Gene knockout

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics

Cite this

Xiang, Y. G., Sun, X., Wang, L., Yan, L. Z., Yang, H., Liu, W., ... Wang, Z. G. (2004). Generation of BPOZ gene knock-out mouse model. Progress in Biochemistry and Biophysics, 31(7), 616-621.

Generation of BPOZ gene knock-out mouse model. / Xiang, You Gui; Sun, Xia; Wang, Long; Yan, Lan Zhen; Yang, Hua; Liu, Wei; Xu, Yong; Xu, Guo Jiang; Wang, Yi; Fei, Jian; Fu, Ji Liang; Wang, Zhu Gang.

In: Progress in Biochemistry and Biophysics, Vol. 31, No. 7, 07.2004, p. 616-621.

Research output: Contribution to journalArticle

Xiang, YG, Sun, X, Wang, L, Yan, LZ, Yang, H, Liu, W, Xu, Y, Xu, GJ, Wang, Y, Fei, J, Fu, JL & Wang, ZG 2004, 'Generation of BPOZ gene knock-out mouse model', Progress in Biochemistry and Biophysics, vol. 31, no. 7, pp. 616-621.
Xiang YG, Sun X, Wang L, Yan LZ, Yang H, Liu W et al. Generation of BPOZ gene knock-out mouse model. Progress in Biochemistry and Biophysics. 2004 Jul;31(7):616-621.
Xiang, You Gui ; Sun, Xia ; Wang, Long ; Yan, Lan Zhen ; Yang, Hua ; Liu, Wei ; Xu, Yong ; Xu, Guo Jiang ; Wang, Yi ; Fei, Jian ; Fu, Ji Liang ; Wang, Zhu Gang. / Generation of BPOZ gene knock-out mouse model. In: Progress in Biochemistry and Biophysics. 2004 ; Vol. 31, No. 7. pp. 616-621.
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