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Review
. 1984:80:1-38.

Introduction of genes into the germ line of animals

R L Brinster, R D Palmiter
Review

Introduction of genes into the germ line of animals

R L Brinster et al. Harvey Lect. 1984.
No abstract available

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Figures

Fig. 1
Fig. 1
Microinjection of new genes into a fertilized mouse egg. The upper picture shows a normal egg secured by a large holding pipette. The tip of the smaller pipette is in the male pronucleus. In the lower picture the pronucleus has been swollen by the introduction of a solution containing new genes through the small injection pipette. Egg is approximately 70 μm in diameter. The tip of the injector pipette is 1.5 μm.
Fig. 2
Fig. 2
Structure of plasmid pMK. (A) Plasmid pMT-TK contains a 3.8 kb genomic Eco R1 fragment that includes the MT-1 gene inserted into the Eco R1 site of pBR322 and the 3.5 kb Barn H1 fragment of the herpes simplex virus type I containing the thymidine kinase (TK) gene inserted into the Barn H1 site of pBR322. The two genes are in the same transcriptional orientation, as shown by the arrows. (B) The fusion plasmid, pMK, was created by digestion of pMT-TK with Bgl II endonuclease followed by ligation with T4 DNA ligase to join the 5′ region of MT-1 to the TK structural gene. The MT mRNA coding sequences are indicated by the large solid boxes, and the TK mRNA coding sequences by the large open boxes. Nontranscribed and intron regions of the two genes are shown by a single line. The pBR322 sequences are indicated by the narrow solid boxes. Restriction sites relevant to the construction are indicated. Also shown are the locations of the TATAA promoter sequences and ATG translation start codons for the two genes (Brinster et al., 1981b; Palmiter et al., 1982a).
Fig. 3
Fig. 3
Diagram of the technique used in making and studying transgenic mice. For detailed procedures see Brinster et al. (1985).
Fig. 4
Fig. 4
Fusion genes redirect gene expression in transgenic mice. The natural genes: human growth hormone, mouse metallothionein-I, rat elastase-1, and SV40 T antigen were expressed in the tissues indicated to the right. When their controlling elements (promoter/enhancer) were fused to different structural genes, then the expression of the structural gene was redirected as dictated by the promoter/enhancer specificity. The solid boxes represent exons, the open boxes introns, the line represents the 5′ flanking sequences including the promoter. The solid symbols in the promoter represent the enhancer elements; there are four functional metal regulatory elements in the mouse metallothionein promoter (circles), probably only one cell-specific enhancer element in the elastase promoter (hexagon), and two 72 bp repeats of the SV40 enhancer (squares). All fusions were made between the cap site (+ 1) and the initiation codon of the respective genes. The genes are not drawn to scale.
Fig. 5
Fig. 5
Expression of a microinjected gene in mice resulted in accelerated growth rate and increased size. The gene consisted of the mouse metallothionein promoter/regulator fused to the rat growth hormone structural gene. These mice were ten weeks old when the picture was taken. The large mouse weighed 44 grams. The small mouse was a litter mate control and weighed 30 grams. For details see Palmiter et al. (1982b).
Fig. 6
Fig. 6
Growth hormone (GH) cascade. The hypothalamus produces the neuropeptides somatostatin (SS) and growth hormone releasing factor (GRF) that inhibit and stimulate, respectively, the release of GH. Growth hormone stimulates hepatocytes to produce insulinlike growth factor I (IGF-I) which acts on peripheral tissues to stimulate growth. Growth hormone also may stimulate peripheral tissues directly. Growth hormone and IGF-I are believed to influence the cascade by a negative feedback effect on the hypothalamus and pituitary. Metallothionein fusion genes are expressed in the liver and several other tissues. Therefore, their products are not controlled by the normal endogenous regulatory mechanism. [MT] indicates the MT promoter was responsible for production of the hormone. The origin of the hormone gene was rat (r), human (h), or bovine (b). (+) and (−) indicate positive and negative influences, respectively.
Fig. 7
Fig. 7
Immunofluorescent localization of human growth hormone in pancreas from a transgenic mouse containing a rat elastase-human growth hormone fusion gene (see Fig. 4). The exocrine acinar cells fluoresce brightly while the endocrine cells (two islets) are dark. For details see Ornitz et al. (1985).
Fig. 8
Fig. 8
Transgenic mouse containing a rat elastase-simian virus 40 T-antigen fusion gene (see Fig. 4). The left picture shows the large abdominal swelling seen in advanced stages of tumor development. The right picture shows two large masses (arrows) which were found on histological examination to be adenocarcinomas of the exocrine pancreas.
Fig. 9
Fig. 9
Transgenic mouse containing a c-myc gene isolated from a mouse plasmacytoma and reintroduced into the germ line by microinjection. In left picture note the greatly enlarged axillary lymph nodes. Right picture demonstrates that most or all lymph nodes (arrows) are enlarged. The tumor phenotype is inherited.
See this image and copyright information in PMC

References

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