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| Construction of Human SP-B Protein Transgenic Mice and Bacterial Pneumonia Model |
| GE Lin1, LIU Xin-yu2, WANG Guirong3 |
1. Tianjin Medical University, Department of Biochemistry and Molecular Biology, Tianjin 300070, China;
2. Key Laboratory of Hormones and Development Ministry of Health, Tianjin Key Laboratory of Metabolic Diseases, Tianjin Metabolic Diseases Hospital & Tianjin Institute of Endocrinology, Tianjin Medical University, Tianjin 300070, China;
3. SUNY Upstate Medical University, Department of Surgery, Syracuse NY 13210, USA |
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Abstract Objective:to construct the transgenic mice expressing human SP-B gene with different alleles in +1580 SNP and the bacterial pneumonia model. Methods:The hSP-B gene was integrated into the mouse chromosome to obtain the F0 generation mice by microinjection technology. The mice were mated with mSP-B gene knockout mice to eliminate the mSP-B gene gradually. Using PCR technique to identify the genotype of mice, and to determine the allele of +1580 locus by sequencing. The Pseudomonas aeruginosa was inoculated into the lung of mice to make the model of bacterial pneumonia, and the control group was injected with the same amount of sterile saline. Results:F2 mice expressed human SP-B protein only, and the expression level of SP-B protein was similar to that in human lung. The +1580 locus alleles T was found in 3 mice families and allele C was found in 1 mice family. After infection for 24 hours, the inflammatory exudation in the alveolar of mice was obvious. A large number of neutrophils were observed in mice alveolus. SP-B protein level was significantly reduced, but there was no significant difference between the different alleles. Conclusions:the construction of human SP-B protein expression transgenic mice model and bacterial pneumonia model were successful. The mice will be a powerful tool for the further studies on physiological function of human SP-B protein and the relationship between +1580 gene polymorphism and lung diseases.
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Received: 11 April 2017
Published: 25 October 2017
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