显微注射法建成HLA-B_(27)转基因大鼠

Establishment of HLA-B_(27) gene transgcnic rats

目的构建带ＨＬＡ－Ｂ_２７基因的转基因大鼠，进一步研究ＨＬＡ－Ｂ_２７与强直性脊柱炎（ＡＳ）的发病原因及其机理。方法应用转基因技术，把已克隆的人类ＡＳ基因ＨＬＡ－Ｂ_２７基因组ＤＮＡ导人Ｗｉｓｔａｒ大鼠受精卵中，并移植到假孕母鼠输卵管内，２１ｄ后，出生２７只原代大鼠。结果经过ＰＣＲ检测阳性后，再继续进行Ｓｏｕｔｈｅｒｎ印迹分析，确认３只大鼠为带ＨＬＡ－Ｂ_２７基因转基因大鼠，整合率为１１．１１％，其中１只转基因大鼠出现外周关节炎及脊柱强直表现。结论本研究进一步表明ＨＬＡ－Ｂ_２７与ＡＳ的强相关性，为研究ＡＳ病因病理及治疗打下了基础。

Objective To generate transgenic rats carrying HLA-B_27 gene, and study the relationship between the HLA-B_27 gene andthe pathogenesis of ankylosing spondylitis (AS). Methods The cloned genomic DNA of human AS gene, HLA-B_27 was injected bymicroinjection technique into the fertilized ovum of Wistar rats, which was then transplanted into the oviduct of pseudopregnantfemale rats. 27 transgenic rats (first generation) were born ailer 21 days. Results in these 27 rats, only 3 carried HLA-B_27 gene.confirmed by PCR and Southern blotting technique. The integrity rate was 1 1. 11% (3/27). Of these transgenic rats, only one ratshowed peripheral arthritis and manitbstations of AS. Conclusion This paper suggested that HLA-B_27 gene has evident correlationwith AS and this study has laid a foundation for the further study of the etiology, pathology and therapy of AS.