本文選題：太子參 + 肌動(dòng)蛋白��； 參考：《分子植物育種》2017年02期

【摘要】：根據(jù)太子參肌動(dòng)蛋白基因PhACT2的已知片段設(shè)計(jì)特異引物,采用RACE技術(shù)擴(kuò)增全長(zhǎng)cDNA,并通過(guò)生物信息學(xué)軟件對(duì)該基因進(jìn)行結(jié)構(gòu)分析。通過(guò)測(cè)序及序列拼接獲得PhACT2全長(zhǎng)cDNA序列,開(kāi)放閱讀框?yàn)? 134 bp,編碼377個(gè)氨基酸殘基,分子量為41.79 k D,理論等電點(diǎn)為5.30。與其他植物同源序列進(jìn)行分析表明,其核苷酸序列與甜菜的同源性較高為88%,氨基酸序列的同源性均在95%以上。進(jìn)化分析表明,PhACT2與擬南芥At ACT7的氨基酸序列僅存在4個(gè)特異位點(diǎn)的氨基酸差異。實(shí)時(shí)熒光定量PCR分析結(jié)果顯示,PhACT2在太子參的不同器官、組織中具有穩(wěn)定的表達(dá),可作為內(nèi)參基因。PhACT2全長(zhǎng)cDNA序列的成功克隆和生物信息學(xué)分析為其在分子生物學(xué)領(lǐng)域的應(yīng)用奠定基礎(chǔ)。
[Abstract]:A specific primer was designed according to the known PhACT2 fragment of the actin gene of Pseudostellaria heterophylla. The full-length cDNA was amplified by RACE and the structure of the gene was analyzed by bioinformatics software.The full-length PhACT2 cDNA sequence was obtained by sequencing and sequence splicing. The open reading frame was 1 134 BP, encoding 377 amino acid residues with a molecular weight of 41.79 KD and a theoretical isoelectric point of 5.30.The results of homology analysis with other plants showed that the homology of nucleotide sequence with sugar beet was 888.The homology of amino acid sequence was more than 95%.Evolutionary analysis showed that there were only four amino acid differences in the amino acid sequence of ACT7 between Ph and Arabidopsis thaliana.The results of real-time fluorescence quantitative PCR analysis showed that PhACT2 was stably expressed in different organs and tissues of Pseudostellaria heterophylla.It can be used as the successful cloning and bioinformatics analysis of the full-length cDNA sequence of the internal reference gene .PhACT2, which lays the foundation for its application in the field of molecular biology.
【作者單位】： 貴陽(yáng)中醫(yī)學(xué)院;
【基金】：國(guó)家自然科學(xué)基金(81460579);國(guó)家自然科學(xué)基金(81160501) 貴州省研究生工作站建設(shè)項(xiàng)目(黔教研合JYSZ字[2014]016)共同資助
【分類(lèi)號(hào)】：S567.53;Q943.2
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本文編號(hào)：1742621