本文選題：XLRS　切入點：視網(wǎng)膜劈裂　出處：《南京醫(yī)科大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：背景和目的：X染色體連鎖視網(wǎng)膜劈裂（X-Linked Retinoschisis，XLRS），又稱先天性視網(wǎng)膜劈裂，主要發(fā)生于男性青少年，是男性青少年黃斑變性的一個主要病因，以視網(wǎng)膜神經(jīng)纖維層撕裂為特征，嚴(yán)重影響視力。該病主要是X染色體連鎖遺傳，基因XLRS1(X-Linked Retinoschisin1,X染色體連鎖視網(wǎng)膜劈裂蛋白1，NCBI Gene ID:6247)的突變經(jīng)由從攜帶者女性經(jīng)由X染色體傳給男性后代而致病，也有少部分病例是常染色體顯性遺傳。本文通過對于一個XLRS病例的研究，發(fā)現(xiàn)了一個新的XLRS1突變位點，，并對該突變位點的致病機(jī)制做了進(jìn)一步的研究，試圖闡明該突變位點的分子致病機(jī)制。 方法：通過視力、眼底鏡檢查、OCT檢查和ERG檢查臨床診斷一例XLRS受累的7周歲男孩。通過抽提該男孩及其父母血液DNA，分別測序XLRS16個外顯子序列，以確定突變位點和致病基因的遺傳模式。后續(xù)研究，通過慢病毒穩(wěn)定轉(zhuǎn)染、融合蛋白技術(shù)和定點突變技術(shù)，建立了能夠穩(wěn)定表達(dá)帶有標(biāo)簽融合蛋白3×FLAG的RS1（Retinoschisin1,視網(wǎng)膜劈裂蛋白）的293T細(xì)胞株。對該細(xì)胞株的RS1蛋白表達(dá)模式以及蛋白的多聚體形成模式進(jìn)行分析。 結(jié)果：確定該XLRS受累男孩病例的XLRS1基因發(fā)生了點突變，突變位點位于CDS區(qū)第332，堿基從野生型的C突變?yōu)門（c.332CT）�；颊吣赣H測序顯示在同樣位點同時存在野生型C和突變型T兩個等位基因�；颊吒赣HXLRS1為野生型。都過嘌呤霉素篩選，獲得了成功表達(dá)RS1野生型和RS1突變型蛋白的兩株293T。蛋白質(zhì)表達(dá)模式分析發(fā)現(xiàn)c.332CT的RS1突變無法分泌至細(xì)胞外。 結(jié)論：發(fā)現(xiàn)了XLRS1基因一個新的突變位點c.332CT，該突變型基因符合X染色體連鎖隱性遺傳模式，可能是突變RS1蛋白的分泌障礙導(dǎo)致了XLRS。
[Abstract]:Background and objective: X chromosome linked retinal cleavage X-Linked Retinoschiosis (XLRSs), also known as congenital retinoschisis, mainly occurs in male adolescents and is a major cause of male adolescent macular degeneration, characterized by retinal nerve fiber layer tear. The disease is mainly X-linked, and the mutation in the gene XLRS1(X-Linked Retinoschisin 1 X linked retinal cleavage protein (1 NCBI Gene ID: 6247) is caused by transmission from the female carrier to the male offspring via the X chromosome. A few cases are autosomal dominant inheritance. In this paper, a new XLRS1 mutation site was found in a XLRS case, and the pathogenicity of the mutation site was further studied. This paper attempts to elucidate the molecular pathogenesis of the mutation site. Methods: a 7 year old boy with XLRS involvement was clinically diagnosed by Oct and ERG. The XLRS16 exons were sequenced by extracting the DNA from the blood of the boy and his parents. In order to determine the genetic pattern of mutation sites and pathogenicity genes, the following studies were carried out, including stable transfection of lentivirus, fusion protein technique and site-directed mutagenesis, A 293T cell line was established which could stably express RS1 Retinoschisin 1 (retinal splitting protein) with labeled fusion protein 3 脳 FLAG. The expression pattern of RS1 protein and the pattern of protein polymorphic formation were analyzed. Results: a point mutation in the XLRS1 gene was identified in the boy case of XLRS involvement. The mutation locus was located in region 332of CDS, and the base mutation was transformed from wild type C to TX c.332CT.Among the same locus, two alleles of wild type C and mutant T were found in the mother of the patient. The father of the patient, XLRS1, was a wild type. All of them were screened by purine mycin. Two strains of RS1 wild-type and RS1 mutant proteins were obtained. The protein expression pattern analysis showed that the RS1 mutation of c.332CT could not be secreted out of the cell. Conclusion: a new mutation locus c.332CTin XLRS1 gene was found. The mutant gene is in line with the X chromosome linkage recessive genetic pattern, which may be caused by the secretion of mutant RS1 protein.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R774.1

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