本文關(guān)鍵詞： 端粒 年齡 實(shí)時定量PCR　出處：《昆明醫(yī)學(xué)院》2006年碩士論文　論文類型：學(xué)位論文

【摘要】：目的 端粒在細(xì)胞正常生理功能運(yùn)轉(zhuǎn)過程中起重要作用，同一組織端粒的長度變化與年齡呈負(fù)相關(guān)，即隨著年齡的增加，端粒長度均表現(xiàn)為逐漸變短的趨勢，這種變化趨勢表現(xiàn)為組織特異性，即同年齡不同組織細(xì)胞端粒的長度不同(在生殖細(xì)胞中，由于端粒酶的作用，端粒長度保持不變)，而且同一組織細(xì)胞端粒長度隨年齡縮短不均衡，，并且發(fā)現(xiàn)這種不均衡大體分為三個階段，嬰幼兒時期端�？s短趨勢最快，青少年時期減緩，至壯年以后速度有所回升。我們采取不同年齡階段同一組織(外周血)樣本排除組織差異后，利用實(shí)時定量PCR的方法通過T/S的比值檢測相對端粒長度，如果檢測的樣本量足夠大，就有可能建立起端粒長度與年齡的相關(guān)曲線以及方程。 方法 278例外周血樣本用改進(jìn)后的酚、氯仿、異戊醇法提取DNA，采用PCR方法，擴(kuò)增不同年齡階段樣本的端粒片斷，測序檢測確定為目的片段，再用實(shí)時定量PCR檢測端粒相對長度。 結(jié)果 普通PCR擴(kuò)增出長度大約為200～600bp之間的多條端粒片斷，測序結(jié)果顯示為目的片段，實(shí)時定量PCR可以擴(kuò)增端粒片斷，利用實(shí)時定量PCR檢測端粒相對長度的方法可行。 結(jié)論 PCR的方法可以擴(kuò)增端粒片斷，與經(jīng)典的Southern雜交方法相比具有：簡便、快捷、操作容易、實(shí)驗(yàn)室費(fèi)用消耗較低等特點(diǎn)。并且通過本次試驗(yàn)對用實(shí)時定量PCR的方法檢測端粒相對長度，做了部分嘗試，認(rèn)為該方法可行，為后續(xù)試驗(yàn)做好準(zhǔn)備。
[Abstract]:Objective telomere plays an important role in the normal physiological function of cells. The change of telomere length in the same tissue is negatively correlated with age. This trend shows tissue specificity, in which telomeres vary in length at the same age (in germ cells, due to the role of telomerase). Telomere length remained unchanged, and the length of telomere in the same tissue cells decreased unevenly with age. It was found that the imbalance was divided into three stages: telomere shortening was the fastest in infancy and slowed down in adolescence. After reaching adulthood, we used the same tissue (peripheral blood) samples at different ages to exclude the difference of tissue, and measured the relative telomere length by the ratio of T / S by real-time quantitative PCR. If the sample size is large enough, it is possible to establish the correlation curve and equation between telomere length and age. Methods two hundred and eighty-eight peripheral blood samples were extracted by modified phenol, chloroform and isoamyl alcohol method. The telomere fragments were amplified by PCR method and sequenced. The relative length of telomere was detected by real-time quantitative PCR. Results A number of telomere fragments with a length of about 200 ~ 600bp were amplified by ordinary PCR. The results of sequencing showed that telomere fragments could be amplified by real-time quantitative PCR. The method of detecting the relative length of telomere by real-time quantitative PCR was feasible. Conclusion the method of PCR can amplify the telomere fragment. Compared with the classical Southern hybridization method, it is simple, fast and easy to operate. The cost of laboratory is low, and some attempts have been made to detect the relative length of telomere by the method of real-time quantitative PCR, and it is considered that this method is feasible and ready for further test.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：D919

【參考文獻(xiàn)】

相關(guān)期刊論文 前7條

1 彭江紅;端粒、端粒酶與細(xì)胞衰老[J];安徽師大學(xué)報(自然科學(xué)版);1998年04期

2 葛璐璐,劉超,陳曉暉,陳向紅,陶黎陽,趙永忠,陳平雁;人外周血白細(xì)胞端粒DNA長度變化與性別的關(guān)系[J];中國法醫(yī)學(xué)雜志;2004年06期

3 謝長江;端粒及端粒酶與細(xì)胞老化及年齡相關(guān)疾病[J];廣州醫(yī)學(xué)院學(xué)報;2002年02期

4 李長勇,任甫;端粒、端粒酶與衰老及年齡的關(guān)系[J];解剖科學(xué)進(jìn)展;2005年03期

5 邵晉晨;端粒酶與細(xì)胞周期及細(xì)胞凋亡關(guān)系的研究進(jìn)展[J];國外醫(yī)學(xué)(生理、病理科學(xué)與臨床分冊);2002年01期

6 楊善民;自發(fā)性高血壓大鼠動脈組織端粒酶活性研究[J];廈門大學(xué)學(xué)報(自然科學(xué)版);1999年02期

7 林進(jìn);謝玨;錢文斌;;系統(tǒng)性紅斑狼瘡患者外周血CD4~+和CD8~+細(xì)胞端粒長度及端粒酶活性的研究[J];浙江大學(xué)學(xué)報(醫(yī)學(xué)版);2005年06期

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