桑葉f3h基因PCR克隆與分析
發(fā)布時(shí)間:2018-10-30 17:38
【摘要】:目的克隆桑葉f3h基因并進(jìn)行測(cè)序分析。方法選定了該基因的PCR擴(kuò)增的上游引物為GTCCGAGACGAAGACGAG,下游引物為CTTGTACATCTCGGTGTA,PCR反應(yīng)體系為95℃預(yù)變性2min→變性95℃15sec、退火58℃5sec、延伸72℃15sec(30個(gè)循環(huán))→延伸72℃2min→4℃結(jié)束。并對(duì)克隆得到的片段進(jìn)行分析。結(jié)果克隆出長(zhǎng)度為515bP的片段,通過(guò)NCBI網(wǎng)站的ORF Finder找到開(kāi)放式閱讀框?yàn)?45bp,翻譯成氨基酸序列為115個(gè)氨基酸,理論分子質(zhì)量為13202.2,等電點(diǎn)為5.43。結(jié)果證明本實(shí)驗(yàn)所擴(kuò)增的基因片段為f3h。結(jié)論為進(jìn)一步克隆f3h全長(zhǎng)基因及下一步研究該基因的表達(dá)調(diào)控打下基礎(chǔ)。
[Abstract]:Objective to clone the F 3 h gene from mulberry leaves and analyze it by sequencing. Methods the upstream primer of PCR amplification of the gene was selected as GTCCGAGACGAAGACGAG, downstream primer as CTTGTACATCTCGGTGTA,PCR reaction system, 95 鈩,
本文編號(hào):2300705
[Abstract]:Objective to clone the F 3 h gene from mulberry leaves and analyze it by sequencing. Methods the upstream primer of PCR amplification of the gene was selected as GTCCGAGACGAAGACGAG, downstream primer as CTTGTACATCTCGGTGTA,PCR reaction system, 95 鈩,
本文編號(hào):2300705
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