2004Zhongguo shiyan dongwuxue zazhiRequires access

Breeding and Identifying of the Homozygous hApoE7 Transgenic Mice

Zuhe Qi

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Abstract

Objective Apolipoprotein E (apoE) is an important serum protein that plays a role in lipid metabolism. Moreover, apoE4 has been identified as a major risk factor for Alzheimer's disease (AD). In order to study the relationship between the gene dose and effect at the whole body level, we bred the homozygous hApoE7 transgenic mice. Methods F1 generation of the C 57BL/6-hApoE7 transgenic mice was mated, using PCR method and Southern blot to test the offspring.The image of the blot was done with laser scanning to identify the copy number of hApoE7, the copy number of the homozygote should be as twice as heterozygote. Then those homozygous mice were cross-bred with normal C 57BL/6 mice to test further. Results 6 F1 generation and 7 F2 generation transgenic mice including 3 homozygotes were identified by Southern blot. The offspring of the crossbreeding of the homozygous mice with normal C 57BL/6 mice were all positive by PCR examination. Conclusion Through breeding the homozygous hApoE7 transgenic mice, we found apoE7 gene could be inherited stably in the transgenic mice and the inheritance pattern of the transgene was compatible with a single autosomal integration site.

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Objective Apolipoprotein E (apoE) is an important serum protein that plays a role in lipid metabolism. Moreover, apoE4 has been identified as a major risk factor for Alzheimer's disease (AD). In order to study the relationship between the gene dose and effect at the whole body level, we bred the homozygous hApoE7 transgenic mice. Methods F1 generation of the C 57BL/6-hApoE7 transgenic mice was mated, using PCR method and Southern blot to test the offspring.The image of the blot was done with laser scanning to identify the copy number of hApoE7, the copy number of the homozygote should be as twice as heterozygote. Then those homozygous mice were cross-bred with normal C 57BL/6 mice to test further. Results 6 F1 generation and 7 F2 generation transgenic mice including 3 homozygotes were identified by Southern blot. The offspring of the crossbreeding of the homozygous mice with normal C 57BL/6 mice were all positive by PCR examination. Conclusion Through breeding the homozygous hApoE7 transgenic mice, we found apoE7 gene could be inherited stably in the transgenic mice and the inheritance pattern of the transgene was compatible with a single autosomal integration site.

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Available abstract

Objective Apolipoprotein E (apoE) is an important serum protein that plays a role in lipid metabolism. Moreover, apoE4 has been identified as a major risk factor for Alzheimer's disease (AD). In order to study the relationship between the gene dose and effect at the whole body level, we bred the homozygous hApoE7 transgenic mice. Methods F1 generation of the C 57BL/6-hApoE7 transgenic mice was mated, using PCR method and Southern blot to test the offspring.The image of the blot was done with laser scanning to identify the copy number of hApoE7, the copy number of the homozygote should be as twice as heterozygote. Then those homozygous mice were cross-bred with normal C 57BL/6 mice to test further. Results 6 F1 generation and 7 F2 generation transgenic mice including 3 homozygotes were identified by Southern blot. The offspring of the crossbreeding of the homozygous mice with normal C 57BL/6 mice were all positive by PCR examination. Conclusion Through breeding the homozygous hApoE7 transgenic mice, we found apoE7 gene could be inherited stably in the transgenic mice and the inheritance pattern of the transgene was compatible with a single autosomal integration site.

Key concepts: Offspring, Transgene, Biology, Genetically modified mouse, Southern blot, Gene, Western blot, Apolipoprotein B

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