關(guān)鍵詞:視網(wǎng)膜變性;基因擴增;序列分析;動物,實驗;聚合酶鏈反應(yīng)
【摘要】 目的 克隆大鼠視網(wǎng)膜緩慢變性/盤膜邊緣蛋白(retinaldegeneration slow/peripherin,RDS/peripherin) 基因。 方法 以SD大鼠的視網(wǎng)膜polyA+ RNA為模版,采用RT-PCR的方法擴增一段約1 555 bp的cDNA片段,并亞克隆至pBluescriptⅡKS(+)載體中,進(jìn)行限制性內(nèi)切酶酶切和序列分析。 結(jié)果 證實所克隆的片段是大鼠的RDS/peripherin cDNA,與Begy等報道的序列[1] 相比,除3′端非翻譯區(qū)的第1 242位的堿基A→G,第1 409~1 411位堿基CA→CCA外,其它序列基本一致[1] 。 結(jié)論 已獲得大鼠RDS/peripherin基因的cDNA,為研究RDS/peripherin基因的功能及在視網(wǎng)膜變性疾病中的作用奠定實驗基礎(chǔ)。
中國圖書資料分類法分類號 R774.13 R730.43 R392.11
Cloning of rat RDS/peripherin gene
GAO Ling,YAN Mi,CHEN Danian,et al.Department of Ophthalmology,The First ClinicalMedical College,West China University of Medical Sciences,Chengdu 610041,China
【Abstract】 Objective Molecularcloning of rat retinal degeneration slow(RDS)gene cDNA. Methods UsingPoly A+ RNA from retina of SD rat as template,a 1 555bp positive cDNAband was obtained by RT-PCR and subcloned into pBluescriptⅡKS(+) vector.The clonedfragment was analyzed with restriction endonucleases and sequencing. Results Ithad been proved that the cloned fragment was rat RDS/peripherin cDNA.Except for thesubstitute of A1 242G and CA 1409-1 411CCA,the other sequences corresponded to that reportedby Begy. Conclusion Rat RDS/peripherin cDNA was obtained.Researcheson function of rat RDS/peripherin gene and its role in retinal degeneration are underway.
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