本文關(guān)鍵詞：重組人腦鈉素在畢赤酵母中的分泌表達(dá)　出處：《吉林大學(xué)》2007年碩士論文　論文類型：學(xué)位論文

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【摘要】： 腦鈉素(Brain Natriuretic Peptide, BNP)又稱B型利鈉肽(B-type Natriuretic Peptide),是利鈉肽(Natriuretic Peptide, NP)家族成員之一,廣泛存在于人體的腦、脊髓、血液、心臟、肺臟和腎臟等多種器官組織中,且主要由心室分泌。研究表明,BNP具有很強(qiáng)的利鈉、利尿、舒張血管和降低血壓等生理功能,在維持細(xì)胞滲透壓、機(jī)體水鹽代謝平衡和血壓穩(wěn)定等方面發(fā)揮著重要作用。BNP濃度的改變?cè)诟哐獕骸⑿哪X血管病和腎病等許多疾病中具有重要意義。血漿BNP水平的檢測(cè),對(duì)于多種心血管疾病的診斷、鑒別診斷、預(yù)后判斷甚至指導(dǎo)治療均有重要價(jià)值。而且BNP已被批準(zhǔn)用于治療急性失代償性心力衰竭,多種拓展適應(yīng)癥也正在進(jìn)行臨床實(shí)驗(yàn)。上述這些事實(shí)提示,BNP具有良好的應(yīng)用前景,大量獲得這一激素,對(duì)心血管疾病和腎臟疾病的診斷、治療以及防治等具有重要意義。 本研究根據(jù)畢赤酵母偏愛(ài)密碼子化學(xué)合成人BNP(hBNP)全基因,以pPICZαC為表達(dá)載體,成功構(gòu)建了重組表達(dá)質(zhì)粒pPICZαC-hBNP,經(jīng)線性化后,電擊法轉(zhuǎn)化畢赤酵母X-33,通過(guò)PCR法篩選出陽(yáng)性轉(zhuǎn)化菌株,從而成功構(gòu)建hBNP畢赤酵母真核表達(dá)體系。陽(yáng)性菌株經(jīng)甲醇誘導(dǎo)培養(yǎng),發(fā)酵上清經(jīng)SDS-PAGE鑒定證明,成功表達(dá)了人腦鈉素,篩選出穩(wěn)定分泌重組人腦鈉素(rhBNP)的酵母菌株,為近一步進(jìn)行rhBNP的大規(guī)模發(fā)酵生產(chǎn)及分離純化奠定了基礎(chǔ)。
[Abstract]:Brain natriuretic peptide (Brain Natriuretic, Peptide, BNP) also called B type natriuretic peptide (B-type Natriuretic Peptide), atrial natriuretic peptide (Natriuretic Peptide, NP is one of the members of the family), exists widely in human brain, spinal cord, blood, heart, lung and kidney and other organ tissues, and secreted by ventricular studies. BNP, has strong natriuretic, diuretic, vasodilator and reduce the blood pressure and other physiological functions in the maintenance of cell osmotic pressure, body water and salt metabolism balance and blood pressure stability plays an important role in the change of.BNP concentration in high blood pressure, plays an important role in many diseases of cardiovascular and cerebrovascular disease and nephropathy in plasma. The level of BNP, a variety of cardiovascular diseases for diagnosis, differential diagnosis, prognosis and guiding treatment has important value. And BNP has been approved for the treatment of acute decompensated heart failure, a variety of indications are expanding In clinical trials, these facts indicate that BNP has good application prospects, and this hormone is of great significance for the diagnosis, treatment and prevention of cardiovascular diseases and kidney diseases.
According to the preferred codon of Pichia pastoris was chemically synthesized BNP (hBNP) gene, expression vector with pPICZ alpha C, successfully constructed the recombinant expression plasmid of pPICZ alpha C-hBNP, after linearization, transformed into Pichia pastoris X-33 by electroporation, screened positive transformed strains by PCR method, so as to build the hBNP Bi pastoris eukaryotic expression system. The positive strain was induced by methanol culture supernatant by SDS-PAGE showed that the expression of brain natriuretic peptide and screened stable secretion of recombinant human brain natriuretic peptide (rhBNP) of the purified yeast strain, laid the foundation for large scale fermentation and separation for further rhBNP.

【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】：2007
【分類號(hào)】：R346

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本文編號(hào)：1379823