人SP-B蛋白转基因小鼠及细菌性肺炎模型的构建

发布时间：2019-01-12 12:28

【摘要】：目的:构建携带人SP-B蛋白+1580 SNP不同等位基因的转基因小鼠并进行细菌性肺炎模型的造模。方法:利用受精卵原核注射技术将hSP-B基因整合至小鼠染色体上获得F0代小鼠,将其与mSP-B基因敲除鼠进行交配,逐步去除转基因小鼠体内m SP-B基因。利用PCR技术鉴定小鼠基因型,通过测序确定+1580位点的等位基因。将铜绿假单胞菌经支气管灌注接种至小鼠肺内进行细菌性肺炎造模,对照组注射等量灭菌生理盐水。结果:F2代小鼠只表达人SP-B蛋白而不表达鼠SP-B蛋白,蛋白表达量与人肺内含量相近,即为构建成功的转基因小鼠。3个小鼠家系+1580位点等位基因为T,1个家系为C。细菌接种(1×10~6CFU/mouse)后24小时,小鼠肺泡内炎症渗出明显,大量中性粒细胞浸润,SP-B蛋白含量明显降低,但不同等位基因间在此条件下无明显差异。结果:成功构建只表达人SP-B蛋白的转基因小鼠模型,细菌性肺炎模型造模成功,为今后进一步研究人SP-B蛋白的生理功能及+1580基因多态性与肺疾病的关系提供了有力的工具。
[Abstract]:Aim: to construct transgenic mice carrying different alleles of human SP-B protein 1580 SNP and model the bacterial pneumonia model. Methods: the hSP-B gene was integrated into the chromosomes of mice by prokaryotic injection of fertilized eggs to obtain F0 generation mice. The mice were mated with mSP-B knockout mice, and the m SP-B gene was gradually removed in transgenic mice. The genotypes of mice were identified by PCR and the alleles at locus 1580 were determined by sequencing. Pseudomonas aeruginosa was infused into the lungs of mice to model bacterial pneumonia, and the control group was injected with the same amount of sterilized saline. Results: F2 mice expressed only human SP-B protein but not mouse SP-B protein. The expression of the protein was similar to that of human lung tissue, that is to say, the transgenic mice were constructed successfully. The allele of 1580 locus was T in three families of mice and one family was C. 24 hours after inoculation (1 脳 10~6CFU/mouse), the alveolar inflammation exudated obviously, a large number of neutrophils infiltrated, and the content of SP-B protein decreased significantly in mice, but there was no significant difference between different alleles under this condition. Results: the transgenic mouse model expressing only human SP-B protein was successfully constructed, and the bacterial pneumonia model was successfully established. It provides a powerful tool to further study the physiological function of human SP-B protein and the relationship between 1580 gene polymorphism and lung diseases.
【作者单位】：天津医科大学生物化学与分子生物学系;天津医科大学代谢病医院内分泌研究所卫生部激素与发育重点实验室天津市代谢性疾病重点实验室;美国纽约州立大学上州医科大学;
【基金】：NIH基金HL096007 迈克尔·康诺利捐赠基金亨德瑞基金天津医科大学科学基金2013KY04资助完成
【分类号】：R-332;R563.1

【相似文献】