SLC34A2基因真核表達(dá)載體的構(gòu)建及其轉(zhuǎn)染人肺泡上皮細(xì)胞的實(shí)驗(yàn)研究
發(fā)布時(shí)間:2018-01-20 15:53
本文關(guān)鍵詞: SLC34A2基因 肺泡上皮細(xì)胞 鈣、磷 肺泡微石癥 出處:《吉林大學(xué)》2008年博士論文 論文類型:學(xué)位論文
【摘要】: SLC34A2是編碼磷酸鈉協(xié)同轉(zhuǎn)運(yùn)蛋白的基因,主要參與無(wú)機(jī)磷代謝,表達(dá)在人體的多種組織中,以肺組織表達(dá)最高。本研究①?gòu)恼H朔谓M織中獲取了該基因完整的cDNA,構(gòu)建了真核表達(dá)載體;②然后將該真核表達(dá)載體用脂質(zhì)體的方法成功轉(zhuǎn)染人肺泡上皮細(xì)胞,觀察在過(guò)表達(dá)的情況下和不同藥物刺激下,細(xì)胞外液中鈣、磷的變化;③并在國(guó)內(nèi)首次對(duì)基因SLC34A2的7個(gè)外顯子進(jìn)行了測(cè)序。④本研究第一次成功克隆了SLC34A2基因,并轉(zhuǎn)染肺泡上皮細(xì)胞;⑤觀察了不同情況下,該基因?qū)?xì)胞外液鈣、磷轉(zhuǎn)運(yùn)的影響;⑥同時(shí)我們?cè)谂R床上跟蹤、治療一肺泡微石癥家系,獲得了肺泡微石癥的第一手臨床資料。本研究填補(bǔ)了國(guó)內(nèi)對(duì)SLC34A2基因研究的空白,是肺泡微石癥等微小結(jié)石類疾病發(fā)病機(jī)制的又一個(gè)途徑,為治療這類疾病提供了可參考的理論價(jià)值,為更深入探討SLC34A2基因的功能和信號(hào)轉(zhuǎn)導(dǎo)機(jī)制奠定了重要的實(shí)驗(yàn)基礎(chǔ)。
[Abstract]:SLC34A2 is a gene encoding sodium phosphate cotransporter. It is mainly involved in inorganic phosphorus metabolism and is expressed in a variety of human tissues. In this study, complete cDNAs were obtained from normal human lung tissue and eukaryotic expression vector was constructed. (2) the eukaryotic expression vector was successfully transfected into human alveolar epithelial cells by liposome, and the changes of calcium and phosphorus in extracellular fluid were observed under the condition of overexpression and under the stimulation of different drugs. (3) the 7 exons of gene SLC34A2 were sequenced for the first time in our country. In this study, SLC34A2 gene was cloned successfully and transfected into alveolar epithelial cells for the first time. (5) the effects of the gene on the transport of calcium and phosphorus in extracellular fluid were observed under different conditions. At the same time, we followed up and treated a pedigree with alveolar microlithiasis, and obtained first-hand clinical data of alveolar microlithiasis. This study filled the blank of SLC34A2 gene research in China. It is another way of pathogenesis of microlithiasis such as alveolar microlithiasis, which provides a theoretical value for the treatment of these diseases. It lays an important experimental foundation for further study on the function and signal transduction mechanism of SLC34A2 gene.
【學(xué)位授予單位】:吉林大學(xué)
【學(xué)位級(jí)別】:博士
【學(xué)位授予年份】:2008
【分類號(hào)】:R346
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