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ABSTRACT: To study the expression of human clotting factor IX cDNA in transgenic mice, which is an essential work on gene therapy for hemophilia B, 3 recombinant constructions containing different lengths of human clotting factor IX cDNA have been introduced into the cultured cells. All of the recombinant constructions were found to be expressed well in vitro. They were then microinjected into the male pronuclei of the fertilized mouse eggs respectively for generating transgenic mice. Unfortunately, none of them was expressed in any transgenic mice. These results show that the expression of the human clotting factor IX cDNA in the transgenic mice can be determined by cis regulatory element(s). As compared with the results from other related works, it is suggested that the cis regulatory element(s) is resided in the 5'-end non-coding region.
Science in China. Series B, Chemistry, life sciences & earth sciences 08/1995; 38(7):825-33.
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ABSTRACT: Two recombinant genes containing human coagulation factor IX cDNA driven by SV40 early promoter or by mouse MT promoter were introduced into the cultured mouse fabroblasts, respectively. It was found that both of them could be expressed in the cultured cells. Then, the two recombinant genes were microinjected into the male pronuclei of fertillized mouse eggs for generating transgenic mice harbouring the introduced genes, respectively. However, we did not observed that the two genes could be expressed in transgenic mice. These results suggest that the in vivo expression of human coagulation factor IX cDNA be controlled by some cis regulatory element (s).
Acta Genetica Sinica 02/1995; 22(3):161-6.
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ABSTRACT: The expression of the chimeric gene delta KpF in which the promoter region of delta-crystallin gene was replaced with the long terminal repeats (LTR) of Friend spleen focus forming virus (SFFV) was studied in transgenic mice for approaching the regulatory mechanism of the expression of chicken delta-crystallin gene. The recombinant plasmid (p delta KpF) DNA containing delta KpF gene was microinjected into male pronuclei of fertilized mouse eggs. The injected eggs were implanted into the oviducts of pseudopregnant female mice and allowed to develop to term. Subsequently, 35 mice from those eggs were born. The mice were analyzed at 2 weeks of age with respect to gene integration and expression by the method of DNA blot hybridization and immunoenzymologic assay. DNA-DNA hybridization indicated that the genomic DNA from one of the mice retained the sequences that hybridized strongly with the probe, p delta KpF. Enzyme linked immunosorbent assay (ELISA) showed that delta KpF gene was expressed in the lens, but not in the brain, muscle and liver. These results demonstrate that the essential regulatory elements for tissue-specific expression of chicken delta-crystallin gene reside in the sequences downstream from +677bp.
Acta Genetica Sinica 02/1992; 19(1):27-33.
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ABSTRACT: A library of genomic DNA has been constructed in EMBL3 lambda phage, from a Chinese hamster/human lymphocytes somatic cell hybrid carrying human chromosome 11 and 20. Recombinants containing human genomic DNA origin can be isolated from the hybrid cell genomic library by using species-specific probe. 8 single copy fragments have been isolated from 13 recombinants. One of them designated as FD11-1 has been identified on chromosome 11 by hybridized it with hybrid cell clone panel and mapped on chromosome 11p11-q11 by chromosome in situ hybridization. On chromosome 11, 3 linkage groups were reported, which located on 11p15, 11p13 and 11q13 respectively. Therefore, the FD11-1 will supply a new locus on chromosome 11 for linkage analysis. Endonuclease recognizing sites and potential recognizing sites on FD11-1 will guide the further RFLP studies.
Acta Genetica Sinica 02/1990; 17(6):469-75.
