發(fā)布時(shí)間：2018-04-11 11:00

  本文選題：酵母胞嘧啶脫氨酶 + 移植物抗宿主病��； 參考：《吉林大學(xué)》2005年碩士論文

【摘要】：目的:探討轉(zhuǎn)染新型自殺基因YCD 的T 淋巴細(xì)胞其體外功能及對(duì)前體藥物的敏感性,為其防治GVHD 提供理論依據(jù)。 方法:①脂質(zhì)體法轉(zhuǎn)染瞬時(shí)包裝細(xì)胞293T 制備含自殺基因YCD 的逆轉(zhuǎn)錄病毒;②將逆轉(zhuǎn)錄病毒導(dǎo)入T 細(xì)胞,并進(jìn)行篩選、擴(kuò)增,獲得轉(zhuǎn)基因T 細(xì)胞;③應(yīng)用PCR 方法檢測(cè)轉(zhuǎn)基因T 細(xì)胞YCD 基因的表達(dá),ELISA 方法檢測(cè)細(xì)胞因子分泌情況,MTT 法檢測(cè)其殺傷功能及對(duì)前體藥物的敏感性。 結(jié)果:①成功將自殺基因YCD 轉(zhuǎn)入293T 細(xì)胞,基因染率可達(dá)38.2%;②PCR 方法可檢測(cè)到轉(zhuǎn)基因T 細(xì)胞中YCD 基因的表達(dá); ③篩選出的轉(zhuǎn)基因T 細(xì)胞其增殖功能正常,與正常T 細(xì)胞相比,對(duì)K562 細(xì)胞的細(xì)胞毒作用無(wú)改變,細(xì)胞因子分泌功能正常;④體外實(shí)驗(yàn)證實(shí)轉(zhuǎn)基因T 細(xì)胞可被38μmol/L 的前體藥物5-FC 全部殺傷。 結(jié)論:①本實(shí)驗(yàn)利用pMSCV-YCD-Puro 載體,成功的將YCD 基因?qū)肴薚細(xì)胞;②轉(zhuǎn)基因T 細(xì)胞的增殖、細(xì)胞因子分泌功能及細(xì)胞毒作用未受影響;③轉(zhuǎn)基因T 細(xì)胞對(duì)前體藥物敏感,可被高度殺傷。
[Abstract]:Aim: to investigate the function of T lymphocytes transfected with novel suicide gene YCD in vitro and its sensitivity to precursor drugs in order to provide theoretical basis for the prevention and treatment of GVHD.Methods Twain 1 liposome method was used to transfect 293T of transient packaging cells to produce retrovirus 2 containing suicide gene YCD into T cells. The T cells were screened and amplified to obtain transgenic T cells.3The expression of YCD gene in transgenic T cells was detected by PCR method. The cytokine secretion was detected by Elisa. The cytotoxicity and sensitivity to precursor drugs were detected by MTT assay.Results the suicide gene YCD was successfully transferred into 293T cells. The expression of YCD gene in transgenic T cells could be detected by 2PCR, and the proliferative function of the screened transgenic T cells was normal, compared with that of normal T cells.The cytotoxicity of K562 cells was not changed and the cytokine secretion function was normal in vitro. The results showed that the transgenic T cells could be completely killed by 38 渭 mol/L precursor drug 5-FC.Conclusion in this experiment, pMSCV-YCD-Puro vector was used to successfully transfer YCD gene into human T cell line 2 transgenic T cell proliferation, cytokine secretion function and cytotoxicity were not affected.Can be highly killed.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2005
【分類號(hào)】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 江千里,王健民,溫麗敏,江汕,周虹;批量快速測(cè)定法測(cè)定標(biāo)志基因?yàn)镚FP的重組病毒滴度[J];第二軍醫(yī)大學(xué)學(xué)報(bào);2002年09期

2 姜義榮,馬道新;自殺基因在異基因骨髓移植中的應(yīng)用[J];國(guó)外醫(yī)學(xué)(腫瘤學(xué)分冊(cè));2003年04期

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本文編號(hào)：1735712