Melanization in albino mice transformed by introducing cloned mouse tyrosinase gene

Development ◽  
1990 ◽  
Vol 108 (2) ◽  
pp. 223-227 ◽  
Author(s):  
S. Tanaka ◽  
H. Yamamoto ◽  
S. Takeuchi ◽  
T. Takeuchi
Keyword(s):  
Transgenic Mice ◽  
Histological Analysis ◽  
Pigment Production ◽  
Flanking Sequence ◽  
First Report ◽  
Tyrosinase Gene ◽  
Flanking Region ◽  
Albino Mice ◽  
Successful Expression

We introduced a mouse tyrosinase minigene, mg-Tyrs-J, in which the authentic genomic 5′ non-coding flanking sequence was fused to a mouse tyrosinase cDNA, into fertilized egges of albino mice. Of the 25 animals that developed from the injected eggs, four mice exhibited pigmented hair and eyes. Histological analysis of the transgenic mice revealed that the melanogenesis was restricted to hair bulbs and eyes. These results suggest that this minigene encodes active tyrosinase protein and that its 5′ flanking region contains the sequences regulating expression of mouse tyrosinase gene. This is the first report of a successful expression of tyrosinase gene and of pigment production in transgenic mice.

Goblet cell-specific expression mediated by theMUC2 mucin gene promoter in the intestine of transgenic mice

1999 ◽  
Vol 276 (3) ◽  
pp. G666-G676 ◽  
Author(s):  
James R. Gum ◽  
James W. Hicks ◽  
Anne-Marie Gillespie ◽  
Elaine J. Carlson ◽  
Lazlo Kömüves ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Goblet Cell ◽  
Human Growth ◽  
Immunohistochemical Analysis ◽  
Regulatory Elements ◽  
Flanking Sequence ◽  
Specific Expression ◽  
Distal Small Intestine ◽  
Mucin Gene ◽  
Flanking Region

The regulation of MUC2, a major goblet cell mucin gene, was examined by constructing transgenic mice containing bases −2864 to +17 of the human MUC25′-flanking region fused into the 5′-untranslated region of a human growth hormone (hGH) reporter gene. Four of eight transgenic lines expressed reporter. hGH message expression was highest in the distal small intestine, with only one line expressing comparable levels in the colon. This contrasts with endogenous MUC2 expression, which is expressed at its highest levels in the colon. Immunohistochemical analysis indicated that goblet cell-specific expression of reporter begins deep in the crypts, as does endogenous MUC2 gene expression. These results indicate that the MUC2 5′-flanking sequence contains elements sufficient for the appropriate expression of MUC2 in small intestinal goblet cells. Conversely, elements located outside this region appear necessary for efficient colonic expression, implying that the two tissues utilize different regulatory elements. Thus many, but not all, of the elements necessary for MUC2 gene regulation reside between bases −2864 and +17 of the 5′-flanking region.

Transcriptional insulation of the human keratin 18 gene in transgenic mice

1993 ◽  
Vol 13 (4) ◽  
pp. 2214-2223
Author(s):  
N Neznanov ◽  
I S Thorey ◽  
G Ceceña ◽  
R G Oshima
Keyword(s):  
Transgenic Mice ◽  
Thymidine Kinase ◽  
Copy Number ◽  
Thymidine Kinase Gene ◽  
Flanking Sequence ◽  
Kinase Gene ◽  
Tissue Specific ◽  
Dependent Behavior ◽  
Flanking Sequences ◽  
Keratin 18

Expression of the 10-kb human keratin 18 (K18) gene in transgenic mice results in efficient and appropriate tissue-specific expression in a variety of internal epithelial organs, including liver, lung, intestine, kidney, and the ependymal epithelium of brain, but not in spleen, heart, or skeletal muscle. Expression at the RNA level is directly proportional to the number of integrated K18 transgenes. These results indicate that the K18 gene is able to insulate itself both from the commonly observed cis-acting effects of the sites of integration and from the potential complications of duplicated copies of the gene arranged in head-to-tail fashion. To begin to identify the K18 gene sequences responsible for this property of transcriptional insulation, additional transgenic mouse lines containing deletions of either the 5' or 3' distal end of the K18 gene have been characterized. Deletion of 1.5 kb of the distal 5' flanking sequence has no effect upon either the tissue specificity or the copy number-dependent behavior of the transgene. In contrast, deletion of the 3.5-kb 3' flanking sequence of the gene results in the loss of the copy number-dependent behavior of the gene in liver and intestine. However, expression in kidney, lung, and brain remains efficient and copy number dependent in these transgenic mice. Furthermore, herpes simplex virus thymidine kinase gene expression is copy number dependent in transgenic mice when the gene is located between the distal 5'- and 3'-flanking sequences of the K18 gene. Each adult transgenic male expressed the thymidine kinase gene in testes and brain and proportionally to the number of integrated transgenes. We conclude that the characteristic of copy number-dependent expression of the K18 gene is tissue specific because the sequence requirements for transcriptional insulation in adult liver and intestine are different from those for lung and kidney. In addition, the behavior of the transgenic thymidine kinase gene in testes and brain suggests that the property of transcriptional insulation of the K18 gene may be conferred by the distal flanking sequences of the K18 gene and, additionally, may function for other genes.

Tissue-specific expression of human CD4 in transgenic mice

1993 ◽  
Vol 13 (5) ◽  
pp. 2952-2958
Author(s):  
F P Gillespie ◽  
L Doros ◽  
J Vitale ◽  
C Blackwell ◽  
J Gosselin ◽  
...  
Keyword(s):  
T Cell ◽  
Transgenic Mice ◽  
Cell Line ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Specific Expression ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
Flanking Region

The gene for the human CD4 glycoprotein, which serves as the receptor for human immunodeficiency virus type 1, along with approximately 23 kb of sequence upstream of the translational start site, was cloned. The ability of 5' flanking sequences to direct tissue-specific expression was tested in cell culture and in transgenic mice. A 5' flanking region of 6 kb was able to direct transcription of the CD4 gene in NIH 3T3 cells but did not result in detectable expression in the murine T-cell line EL4 or in four lines of transgenic mice. A larger 5' flanking region of approximately 23 kb directed high-level CD4 transcription in the murine T-cell line EL4 and in three independent lines of transgenic mice. Human CD4 expression in all tissues analyzed was tightly correlated with murine CD4 expression; the highest levels of human CD4 RNA expression were found in the thymus and spleen, with relatively low levels detected in other tissues. Expression of human CD4 protein in peripheral blood mononuclear cells was examined by flow cytometry in these transgenic animals and found to be restricted to the murine CD4+ subset of lymphocytes. Human CD4 protein, detected with an anti-human CD4 monoclonal antibody, was present on the surface of 45 to 50% of the peripheral blood mononuclear cells from all transgenic lines.

Phenobarbital responsiveness conferred by the 5′-flanking region of the rat CYP2B2 gene in transgenic mice

Gene ◽  
1999 ◽  
Vol 228 (1-2) ◽  
pp. 169-179 ◽  
Author(s):  
Richard Ramsden ◽  
Nancy B Beck ◽  
Karen M Sommer ◽  
Curtis J Omiecinski
Keyword(s):  
Transgenic Mice ◽  
Flanking Region

Genomic deletions in Rhodococcus based on transformation of linear heterologous DNA

Microbiology ◽  
2021 ◽  
Author(s):  
Divya Singhi ◽  
Shabnam Parwin ◽  
Preeti Srivastava
Keyword(s):  
Rapid Method ◽  
Genome Engineering ◽  
Cost Effective ◽  
Cell Disruption ◽  
Content Type ◽  
First Report ◽  
Genomic Deletions ◽  
Linear Dna ◽  
Link Type ◽  
Successful Expression

Several genome engineering methods have been developed for Rhodococcus . However, they suffer from limitations such as extensive cloning, multiple steps, successful expression of heterologous genes via plasmid etc. Here, we report a rapid method for performing genomic deletions/disruptions in Rhodococcus spp. using heterologous linear DNA. The method is cost effective and less labour intensive. The applicability of the method was demonstrated by successful disruption of rodA and orphan parA. None of the disrupted genes were found to be essential for the viability of the cell. Disruption of orphan parA and rodA resulted in elongated cells and short rods, respectively. This is the first report demonstrating disruption of rodA and orphan parA genes by electroporation of heterologous linear DNA in Rhodococcus spp.

A yeast artificial chromosome covering the tyrosinase gene confers copy number-dependent expression in transgenic mice

Nature ◽  
1993 ◽  
Vol 362 (6417) ◽  
pp. 258-261 ◽  
Author(s):  
Andreas Schedl ◽  
Lluís Montoliu ◽  
Gavin Kelsey ◽  
Günther Schütz
Keyword(s):  
Transgenic Mice ◽  
Copy Number ◽  
Yeast Artificial Chromosome ◽  
Artificial Chromosome ◽  
Tyrosinase Gene

scholarly journals Endogenous Production of n-3 Polyunsaturated Fatty Acids Promotes Fracture Healing in Mice

2017 ◽  
Vol 2017 ◽  
pp. 1-6 ◽  
Author(s):  
Yuhui Chen ◽  
He Cao ◽  
Dawei Sun ◽  
Changxin Lin ◽  
Liang Wang ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Fracture Healing ◽  
Histological Analysis ◽  
Callus Formation ◽  
Healing Process ◽  
Proximal Femoral Fracture ◽  
Fracture Repair ◽  
Micro Ct ◽  
Micro Computed Tomography ◽  
X Ray

Bone fracture is a global healthcare issue for high rates of delayed healing and nonunions. Although n-3 polyunsaturated fatty acid (PUFA) is considered as a beneficial factor for bone metabolism, only few studies till date focused on the effects of n-3 PUFAs on fracture healing. In this study, we investigated the effect of endogenous n-3 PUFAs on fracture healing by measuring femur fracture repair in bothfat-1transgenic mice and WT mice. Proximal femoral fracture model was established infat-1transgenic mice and WT mice, respectively, and then the fracture was analyzed by using X-ray, micro-computed tomography (micro-CT), and histological assessment at 7, 14, 21, 28, and 35 days after fixation. The results showed that compared with WT mice,fat-1mice exhibited acceleration in fracture healing through radiographic and histological analysis (18–21 days versus 21–28 days postfracture). Meanwhile, X-ray and micro-CT analysis that showed better remodeling callus formation were in thefat-1group compared to WT group. Furthermore, histological analysis revealed that endogenous n-3 PUFAs promoted local endochondral ossification and accelerated the remodeling of calcified calluses after fracture. In conclusion, the present study indicated that endogenously produced n-3 PUFAs promote fracture healing process and accelerate bone remodeling in mice, and supplementation of n-3 PUFAs was positively associated with fracture healing.

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