重组人脑钠素在毕赤酵母中的分泌表达

发布时间：2018-01-04 19:34

本文关键词：重组人脑钠素在毕赤酵母中的分泌表达　出处：《吉林大学》2007年硕士论文　论文类型：学位论文

【摘要】： 脑钠素(Brain Natriuretic Peptide, BNP)又称B型利钠肽(B-type Natriuretic Peptide),是利钠肽(Natriuretic Peptide, NP)家族成员之一,广泛存在于人体的脑、脊髓、血液、心脏、肺脏和肾脏等多种器官组织中,且主要由心室分泌。研究表明,BNP具有很强的利钠、利尿、舒张血管和降低血压等生理功能,在维持细胞渗透压、机体水盐代谢平衡和血压稳定等方面发挥着重要作用。BNP浓度的改变在高血压、心脑血管病和肾病等许多疾病中具有重要意义。血浆BNP水平的检测,对于多种心血管疾病的诊断、鉴别诊断、预后判断甚至指导治疗均有重要价值。而且BNP已被批准用于治疗急性失代偿性心力衰竭,多种拓展适应症也正在进行临床实验。上述这些事实提示,BNP具有良好的应用前景,大量获得这一激素,对心血管疾病和肾脏疾病的诊断、治疗以及防治等具有重要意义。本研究根据毕赤酵母偏爱密码子化学合成人BNP(hBNP)全基因,以pPICZαC为表达载体,成功构建了重组表达质粒pPICZαC-hBNP,经线性化后,电击法转化毕赤酵母X-33,通过PCR法筛选出阳性转化菌株,从而成功构建hBNP毕赤酵母真核表达体系。阳性菌株经甲醇诱导培养,发酵上清经SDS-PAGE鉴定证明,成功表达了人脑钠素,筛选出稳定分泌重组人脑钠素(rhBNP)的酵母菌株,为近一步进行rhBNP的大规模发酵生产及分离纯化奠定了基础。
[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.

【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R346

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