本文選題：牦牛 + 溶菌酶　； 參考：《黑龍江畜牧獸醫(yī)》2017年09期

【摘要】：為了通過定向進化的方法提高牦牛溶菌酶的活性,試驗選用3個牦�；験SL1a、YSL3c和YML,通過DNA洗牌技術(shù)進行序列重組以及酶活篩選,獲得了一個重組基因HYL,檢測其酶活,構(gòu)建重組子基因HYL的真核表達載體,轉(zhuǎn)化畢赤酵母X33,并對重組HYL溶菌酶的活性進行檢測。結(jié)果表明:HYL與YSL1a、YSL3c和YML的序列相似性分別為85.36%、96.04%和83.11%,突變核苷酸分別為65,17,75個;重組序列與3個親本基因相比均有序列缺失。篩選到一個酶活達667.72 U/mg的陽性克隆,其活性高于親本基因中YSL3c真核表達產(chǎn)物。說明本試驗中牦牛HYL的序列發(fā)生了序列重組,而且其真核表達產(chǎn)物具有活性。
[Abstract]:In order to improve the lysozyme activity of yak by directed evolution, three yak genes YSL1aYSL3c and YMLwere selected and sequenced and screened by DNA shuffling technique. A recombinant gene HYL was obtained and its enzyme activity was detected. The eukaryotic expression vector of recombinant gene HYL was constructed and transformed into Pichia pastoris X33. The lysozyme activity of recombinant HYL was detected. The results showed that the similarity of the sequence of 1: HYL with YSL1aHYSL3c and YML was 85.36% and 83.11%, respectively, and the mutation nucleotides were 65.17 and 75, respectively, and the sequence deletion of the recombinant gene was compared with that of the three parental genes. A positive clone with an enzyme activity of 667.72 U/mg was screened and its activity was higher than that of the eukaryotic expression product of YSL3c in the parent gene. The results showed that the sequence of yak HYL was recombined and its eukaryotic expression product was active.
【作者單位】： 西南民族大學(xué)生命科學(xué)與技術(shù)學(xué)院;貴州省桐仁市農(nóng)業(yè)委員會;
【基金】：四川省青年科技創(chuàng)新團隊項目(2015TD0025) 科技部科技支撐項目(2014BAD13B03) 中央高校基本科研業(yè)務(wù)費專項資金項目(2014NZYQN59)
【分類號】：S823.85

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1 陳偉偉;堿性植酸酶的定向進化及其在不同宿主中的表達[D];浙江大學(xué);2015年

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本文編號：1965443