發(fā)布時(shí)間：2018-05-12 15:55

  本文選題：UAA編碼 + 表達(dá)體系��； 參考：《吉林大學(xué)》2017年碩士論文

【摘要】：與所有的生命過(guò)程一樣,外源基因在原核細(xì)胞中的表達(dá)包括兩個(gè)主要過(guò)程,即DNA轉(zhuǎn)錄成mRNA和mRNA翻譯成蛋白質(zhì)。大腸桿菌一直是基因工程中應(yīng)用最為廣泛的表達(dá)系統(tǒng)。包含非編碼氨基酸的蛋白質(zhì)的過(guò)量表達(dá)已引起越來(lái)越多科學(xué)家的關(guān)注。非編碼氨基酸的定點(diǎn)摻入,使關(guān)于蛋白質(zhì)結(jié)構(gòu)和功能的化學(xué)假定的特異檢測(cè)成為可能。具有新化學(xué)性質(zhì)的非編碼氨基酸的定點(diǎn)摻入,可能使蛋白質(zhì)產(chǎn)生新的功能。本文將pEVOL載體中對(duì)應(yīng)琥珀密碼子UAG的tRNA合酶進(jìn)行定點(diǎn)突變,將其改造成對(duì)應(yīng)赭石密碼子UAA的tRNA/氨酰tRNA合酶表達(dá)系統(tǒng),將UAA密碼子變成了對(duì)應(yīng)8-羥基喹啉丙氨酸的非編碼氨基酸表達(dá)體系,并表達(dá)了摻入8-羥基喹啉丙氨酸的金屬硫蛋白-GFP融合蛋白,驗(yàn)證了該蛋白的金屬結(jié)合能力。
[Abstract]:As in all life processes, the expression of exogenous genes in prokaryotic cells involves two main processes: DNA transcription into mRNA and mRNA translation into proteins. E. coli has been the most widely used expression system in genetic engineering. Overexpression of proteins containing non-encoded amino acids has attracted more and more attention from scientists. The incorporation of non-coding amino acids makes possible the specific detection of chemical assumptions about the structure and function of proteins. The incorporation of non-coding amino acids with new chemical properties may produce new functions for proteins. In this paper, the tRNA synthase corresponding to the amber codon UAG in the pEVOL vector was mutated and transformed into the tRNA/ aminoyl tRNA synthase expression system corresponding to the ochre codon UAA. The UAA codon was transformed into a non-coding amino acid expression system corresponding to 8-hydroxyquinoline-alanine, and the fusion protein of metallothionein GFP was expressed with 8-hydroxyquinoline-alanine, which proved the metal-binding ability of the protein.
【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】：2017
【分類號(hào)】：Q78

【相似文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前2條

1 張晨;UAA編碼氨基酸表達(dá)體系的構(gòu)建[D];吉林大學(xué);2017年

2 陳新駿;人類精子非編碼氨基酸多樣性的研究[D];山東大學(xué);2017年

，

本文編號(hào)：1879222