CAJ | 학술논문

背景:1200009K10基因来源于成年小鼠肺cDNA基因库,于2002-01公布,其功能暂时还未明确.预测其蛋白含有5个区域:Kelch基序,Kelch区,BTB/POZ区,BTB区以及β推进区,以上所有结构均参与蛋白之间的相互作用和一些酶的活性表达.目的:观察1200009K10基因在小鼠视网膜中的表达方式.设计:观察对比实验.单位:中山大学眼科中心研究所.材料:实验用rd小鼠和rds小鼠以及正常对照的C3B小鼠传代小鼠模型购自Jackson实验室(巴港,缅因州04609,美国),所有动物均在中山大学实验动物中心二等动物单元饲养.方法:从小鼠中提取分离视网膜RNA,通过急速冷凝cDNA扩增法扩大1200009K10基因,采用基因特性定量聚合酶链反应对rd小鼠和rds小鼠以及正常对照的C3B小鼠视网膜中该基因的表达进行分析.主要观察指标:3种小鼠视网膜1200009K10基因的表达含量和表达方式.结果:从小鼠视网膜中提取分离的视网膜RNA经过急速cDNA冷凝扩增后,获得了1种与小鼠1200009K10基因序列基本相同的克隆物.小鼠视网膜1200009K10基因的表达分为6个阶段:出生后7,12,25,37,50 d(接近性成熟)及出生后150 d.rds,rd和C3B小鼠视网膜中1200009K10基因表达于小鼠出生后7 d较低,7～12 d较高,12～50 d稳定,最后略有增加.C3B小鼠视网膜中1200009K10基因表达趋势和水平与rds,rd小鼠基本相同,出生12 d后1200009K10基因的表达水平高于rds,rd小鼠.rds,rd和C3B小鼠视网膜1200009K10基因表达水平有所不同,其表达方式在3者之间无明显差异.结论:1200009K10基因可能不参与视网膜变性的发展过程.
배경:1200009K10기인래원우성년소서폐cDNA기인고,우2002-01공포,기공능잠시환미명학.예측기단백함유5개구역:Kelch기서,Kelch구,BTB/POZ구,BTB구이급β추진구,이상소유결구균삼여단백지간적상호작용화일사매적활성표체.목적:관찰1200009K10기인재소서시망막중적표체방식.설계:관찰대비실험.단위:중산대학안과중심연구소.재료:실험용rd소서화rds소서이급정상대조적C3B소서전대소서모형구자Jackson실험실(파항,면인주04609,미국),소유동물균재중산대학실험동물중심이등동물단원사양.방법:종소서중제취분리시망막RNA,통과급속냉응cDNA확증법확대1200009K10기인,채용기인특성정량취합매련반응대rd소서화rds소서이급정상대조적C3B소서시망막중해기인적표체진행분석.주요관찰지표:3충소서시망막1200009K10기인적표체함량화표체방식.결과:종소서시망막중제취분리적시망막RNA경과급속cDNA냉응확증후,획득료1충여소서1200009K10기인서렬기본상동적극륭물.소서시망막1200009K10기인적표체분위6개계단:출생후7,12,25,37,50 d(접근성성숙)급출생후150 d.rds,rd화C3B소서시망막중1200009K10기인표체우소서출생후7 d교저,7～12 d교고,12～50 d은정,최후략유증가.C3B소서시망막중1200009K10기인표체추세화수평여rds,rd소서기본상동,출생12 d후1200009K10기인적표체수평고우rds,rd소서.rds,rd화C3B소서시망막1200009K10기인표체수평유소불동,기표체방식재3자지간무명현차이.결론:1200009K10기인가능불삼여시망막변성적발전과정.
BACKGROUND: 1200009K10 gene is from the lung cDNA pool of adult mouse and announced in January 2002, but its function is still not known.The predicted protein has five domains: Kelch motif, Kelch domain, BTB/POZ domain, BTB domain and β propeller domain, all of which are involved in protein-protein interactions and some enzymatic activities.OBJECTIVE: To explore the correlation between the expression of 1200009K10 gene and retinitis pigmentosa in mice.DESIGN: Observational and comparative trial.SETTING: Institute of Zhongshan Ophthalmic Center of Sun Yat-sen University.MATERIALS: The rd mice, rds mice and passage mice from C3B mice as normal control were purchased from Jackson Laboratory (Bar Harbor,Maine 04609, USA), and raised in second class unit of Experimental Animal Center of Sun Yat-sen University.METHODS: The retinal RNA of mice was extracted and isolated and 1200009K10 gene was amplified by rapid amplification of eDNA ends method (RACE). The gene expressions in the retina of rds, rd and C3B mice were analyzed, respectively by gene-specific real-time quantitative PCR.MAIN OUTCOME MEASURES: The expression contents and patterns of 1200009K10 gene in retina of three kinds of mice.RESULTS: One clone was obtained after RACE of retinal RNA extracted from retina of mice, which had an almost identical sequence with the 1200009K10 gene. Expression of 1200009K10 gene was classified into 6stages: 7, 12, 25, 37, 50 (near sex maturity) and 150 days after born. The expressions of 1200009K10 gene in the retina of rds, rd and C3B mice were low at postnatal day 7 (P7), higher between P7-12, stable between P12-50, and finally increased a little. The expression trend and level of 1200009K10 gene of C3B mice was nearly the same with that of the rds and rd mice, and higher than that of rds and rd mice at P12. Although there were differences in expression levels of 1200009K10 gene among the three kinds of mice, the expression patterns were almost the same.CONCLUSION: 1200009K10 gene may not participate in the development of retinal degeneration.