本文選題：白細(xì)胞介素15 + 克隆　； 參考：《吉林大學(xué)》2014年碩士論文

【摘要】：白細(xì)胞介素15（Interleukin-15,IL-15）是一種多能的細(xì)胞因子，能夠促進(jìn)淋巴細(xì)胞和NK細(xì)胞的增殖且對(duì)免疫系統(tǒng)中各種免疫細(xì)胞都具有重要的調(diào)節(jié)作用。IL-15表現(xiàn)出與IL-2在抗腫瘤、免疫調(diào)節(jié)等方面相似的生物學(xué)作用而備受關(guān)注。 本實(shí)驗(yàn)利用RT-PCR技術(shù)從人外周血單個(gè)核細(xì)胞中克隆了人IL-15cDNA部分序列，并以其為模板分別擴(kuò)增出IL-15編碼序列（coding sequence，CDS）以及引入CD8α跨膜肽的IL-15編碼序列，利用基因組技術(shù)構(gòu)建分泌型人IL-15克隆載體pMD T-IL-15、表達(dá)載體pVITRO2-IL-15以及膜結(jié)合型IL-15克隆載體pMD T-mbIL-15、表達(dá)載體pVITRO2-mbIL-15。分別將表達(dá)載體轉(zhuǎn)染K562細(xì)胞，檢測(cè)分泌型IL-15在K562細(xì)胞中表達(dá)情況，并通過(guò)細(xì)胞流式技術(shù)鑒定，以期獲得能穩(wěn)定表達(dá)的膜結(jié)合IL-15的K562細(xì)胞株。對(duì)進(jìn)一步研究其在免疫系統(tǒng)中的重要作用，并為其在腫瘤基因治療、免疫治療等奠定了基礎(chǔ)。 結(jié)果表明，本實(shí)驗(yàn)成功克隆了人IL-15CDS序列，，在此基礎(chǔ)上正確構(gòu)建了重組人IL-15分泌型真核表達(dá)載體pVITRO2-IL-15和重組人IL-15跨模型真核表達(dá)載體pVITRO2-mbIL-15。最后通過(guò)將重組真核表達(dá)載體分別轉(zhuǎn)染K562細(xì)胞，檢測(cè)了分泌型IL-15在K5625細(xì)胞中的表達(dá)，并獲得穩(wěn)定表達(dá)膜結(jié)合IL-15蛋白的重組K562細(xì)胞株。
[Abstract]:Interleukin 15 Interleukin-15 (IL-15) is a multipotent cytokine that promotes the proliferation of lymphocytes and NK cells and plays an important role in regulating all immune cells in the immune system. IL-15 is associated with IL-2 in anti-tumor activity.Immunomodulation and other similar biological functions have attracted much attention.In this study, we cloned human IL-15cDNA partial sequence from human peripheral blood mononuclear cells by RT-PCR technique, and used it as template to amplify the coding sequence of IL-15 coding sequence sequence sequence and the IL-15 coding sequence that introduced CD8 偽 transmembrane peptide.The secretory human IL-15 clone vector pMD T-IL-15, the expression vector pVITRO2-IL-15 and the membrane-binding IL-15 clone vector pMD T-mbIL-15 were constructed by genomic technique. The expression vector pVITRO2-mbIL-15 was constructed.The expression vectors were transfected into K562 cells, and the expression of secretory IL-15 in K562 cells was detected. The expression of secretory IL-15 in K562 cells was identified by flow cytometry.It can be used to further study its important role in the immune system and lay a foundation for its application in tumor gene therapy and immunotherapy.The results showed that the human IL-15CDS sequence was cloned successfully, and the recombinant human IL-15 secreted eukaryotic expression vector pVITRO2-IL-15 and the recombinant human IL-15 transmodel eukaryotic expression vector pVITRO2-mbIL-15 were constructed.Finally, the recombinant eukaryotic expression vector was transfected into K562 cells, and the expression of secretory IL-15 in K5625 cells was detected, and a recombinant K562 cell line with stable expression of membrane binding IL-15 protein was obtained.
【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】：2014
【分類號(hào)】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 姜云霞;王冰;崔韶暉;趙軼男;楊寶靈;段艷;王秀武;張樹(shù)彪;;陽(yáng)離子脂質(zhì)體基因載體的細(xì)胞轉(zhuǎn)染研究[J];安徽農(nóng)業(yè)科學(xué);2009年17期

2 劉運(yùn)龍;程遠(yuǎn)國(guó);劉學(xué)龍;;干擾素研究進(jìn)展[J];動(dòng)物醫(yī)學(xué)進(jìn)展;2008年02期

3 沈永泉,何維;白細(xì)胞介素-15及其受體[J];國(guó)外醫(yī)學(xué)(分子生物學(xué)分冊(cè));1996年06期

4 張樹(shù)民,洪梅;JAK-STAT信號(hào)轉(zhuǎn)導(dǎo)機(jī)制及其與Ras通路的關(guān)系[J];國(guó)外醫(yī)學(xué)(分子生物學(xué)分冊(cè));1999年04期

5 蘇琴;白細(xì)胞介素-15研究進(jìn)展[J];國(guó)外醫(yī)學(xué)(分子生物學(xué)分冊(cè));2000年02期

6 朱法良;IL-15對(duì)前T細(xì)胞發(fā)育的調(diào)節(jié)作用[J];國(guó)外醫(yī)學(xué)(免疫學(xué)分冊(cè));2000年05期

7 杜肖娜;IL-15與炎癥、感染和腫瘤的關(guān)系[J];國(guó)外醫(yī)學(xué)(免疫學(xué)分冊(cè));2000年06期

8 查紀(jì)坤,舒紅兵;腫瘤壞死因子信號(hào)傳導(dǎo)的分子機(jī)理[J];中國(guó)科學(xué)(C輯:生命科學(xué));2002年01期

9 馬德星,李廣興;白細(xì)胞介素-15研究進(jìn)展[J];吉林畜牧獸醫(yī);2004年07期

10 薛莉,何維;白細(xì)胞介素-15功能最新研究進(jìn)展[J];上海免疫學(xué)雜志;1998年01期

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