發(fā)布時(shí)間：2018-04-29 07:54

  本文選題：陰陽虛證 + 糖皮質(zhì)激素受體��； 參考：《第二軍醫(yī)大學(xué)》2005年碩士論文

【摘要】：目的:構(gòu)建hGRα重組原核表達(dá)質(zhì)粒并進(jìn)行誘導(dǎo)表達(dá),為下一步研制hGRα ELISA檢測試劑盒和進(jìn)一步研究GR與中醫(yī)學(xué)陰陽虛證的關(guān)系服務(wù)。 方法:以含有hGRα cDNA的重組克隆質(zhì)粒pRShGRα為模板,通過PCR擴(kuò)增得到hGRα的兩種基因片段,將這兩種基因片段分別克隆到pGEM-T載體上,進(jìn)而亞克隆到pGEX-4T-3表達(dá)載體上。將這兩種重組表達(dá)質(zhì)粒進(jìn)行酶切、測序鑒定后,分別轉(zhuǎn)化大腸桿菌DH5α和BL21(DE3),并在一定的條件下進(jìn)行重組蛋白的誘導(dǎo)表達(dá),誘導(dǎo)表達(dá)產(chǎn)物行SDS-PAGE和Western blotting分析。 結(jié)果:酶切和測序結(jié)果證明hGRα的兩種基因片段均成功地重組入pGEX-4T-3載體。誘導(dǎo)表達(dá)產(chǎn)物的SDS-PAGE和Western blotting分析均提示有hGRα目的蛋白特異表達(dá)。 結(jié)論:本實(shí)驗(yàn)成功構(gòu)建了hGRα的兩種重組原核表達(dá)質(zhì)粒,并成功誘導(dǎo)了它們的原核表達(dá),為下一階段的研究奠定了基礎(chǔ)。
[Abstract]:Objective: to construct the recombinant prokaryotic expression plasmid of hGR 偽 and express it in order to further study the relationship between gr and yin and yang deficiency syndrome in traditional Chinese medicine. Methods: using the recombinant clone plasmid pRShGR 偽 containing hGR 偽 cDNA as template, two kinds of hGR 偽 gene fragments were amplified by PCR and cloned into pGEM-T vector respectively, and then subcloned into pGEX-4T-3 expression vector. The two recombinant expression plasmids were digested by enzyme and identified by sequencing. The recombinant proteins were transformed into Escherichia coli DH5 偽 and BL21DE-3, respectively. The recombinant protein was induced and expressed under certain conditions. The induced expression products were analyzed by SDS-PAGE and Western blotting. Results: the results of restriction endonuclease digestion and sequencing showed that the two gene fragments of hGR 偽 were successfully recombined into pGEX-4T-3 vector. Both SDS-PAGE and Western blotting analysis of induced expression products indicated that hGR 偽 target protein was specifically expressed. Conclusion: in this experiment, two recombinant prokaryotic expression plasmids of hGR 偽 were successfully constructed, and their prokaryotic expression was induced successfully, which laid a foundation for the next stage of research.
【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2005
【分類號(hào)】：R346

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 凌昌全，李敏，盧軍華，譚金興，張紅武;參附湯對失血性休克大鼠糖皮質(zhì)激素及其受體的影響[J];第二軍醫(yī)大學(xué)學(xué)報(bào);1996年03期

2 李敏，，趙法O，郭俊生，李正銀;加味生脈散對熱損傷大鼠糖皮質(zhì)激素及其受體的影響[J];第二軍醫(yī)大學(xué)學(xué)報(bào);1996年06期

3 徐克川;張善澄;;甲亢類陰虛大鼠肝胞液糖皮質(zhì)激素受體的實(shí)驗(yàn)研究[J];廣州中醫(yī)學(xué)院學(xué)報(bào);1990年03期

4 張斌,徐仁寶;單克隆抗體檢測糖皮質(zhì)激素受體的方法[J];生物化學(xué)與生物物理進(jìn)展;1991年06期

5 施賽珠,沈自尹,陳澤霖,顧天甲,張麗麗,查良倫,黃志昌;腎陽虛病人下丘腦、垂體、腎上腺皮質(zhì)系統(tǒng)功能的觀察[J];上海中醫(yī)藥雜志;1978年01期

6 凌昌全,李敏,朱德增,陳U

本文編號(hào)：1819072