家蚕miR-0001和miR-0015体外下调BmFib-L基因的表达

目的：探究新发现的两个miRNA对家蚕丝素轻链基因BmFib-L的调控作用。创新点：在家蚕后部丝腺中发现两个新的miRNA,并首次证明它们对家蚕丝素蛋白轻链基因BmFib-L有负调控作用。方法：本实验通过生物信息学分析,从家蚕后部丝腺miRNA高通量测序获得的新miRNA中,筛选出两个可能对家蚕丝素蛋白轻链基因BmFib-L有调控作用的miRNA,即bmo-miR-0001和bmo-miR-0015。设计茎环引物,采用反转录聚合酶链反应（RT-PCR）方法对家蚕5龄3 d头部、表皮、脂肪体、马氏管、精巢/卵巢、丝腺（前、中、后）、气管、中肠和血淋巴细胞等12个不同组织的bmo-miR-0001...

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Published in:浙江大学学报：B卷英文版 2016, Vol.17 (2), p.127-135
Main Author: Chen CHEN Yang-yang FAN Xin WANG Fei SONG Tao JIANG Ping QIAN Shun-ming TANG Xing-jia SHEN
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Language:English
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BmFib-L
bmo-miR-0001
bmo-miR-0015
miRNA
功能验证
家蚕
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description 目的：探究新发现的两个miRNA对家蚕丝素轻链基因BmFib-L的调控作用。创新点：在家蚕后部丝腺中发现两个新的miRNA,并首次证明它们对家蚕丝素蛋白轻链基因BmFib-L有负调控作用。方法：本实验通过生物信息学分析,从家蚕后部丝腺miRNA高通量测序获得的新miRNA中,筛选出两个可能对家蚕丝素蛋白轻链基因BmFib-L有调控作用的miRNA,即bmo-miR-0001和bmo-miR-0015。设计茎环引物,采用反转录聚合酶链反应（RT-PCR）方法对家蚕5龄3 d头部、表皮、脂肪体、马氏管、精巢/卵巢、丝腺（前、中、后）、气管、中肠和血淋巴细胞等12个不同组织的bmo-miR-0001和bmo-miR-00015进行半定量表达分析。并采用双荧光报告基因检测系统进一步在细胞水平上验证bmo-miR-0001和bmo-miR-0015对BmFib-L表达的调控作用。结论：本实验中RT-PCR结果显示,这两个新miRNA在家蚕后部丝腺中表达量最高（图4）。双荧光报告基因检测结果显示,报告基因荧光素酶活性明显低于阳性对照组（图7）,转染bmo-miR-0001和bmo-miR-0015表达载体的细胞,报告基因和荧光素酶活性分别只及对照组60%和69%。t检验分析结果显示两个实验组与对照组之间差异都达到显著水平（P〈0.05）。由此可见,bmo-miR-0001和bmo-miR-0015在体外对BmFib-L的表达具有显著的抑制作用。
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bmo-miR-0001 ; bmo-miR-0015 ; miRNA ; 功能验证 ; 家蚕</subject><ispartof>浙江大学学报：B卷英文版, 2016, Vol.17 (2), p.127-135</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://image.cqvip.com/vip1000/qk/86281A/86281A.jpg</thumbnail><link.rule.ids>315,786,790,4043</link.rule.ids></links><search><creatorcontrib>Chen CHEN Yang-yang FAN Xin WANG Fei SONG Tao JIANG Ping QIAN Shun-ming TANG Xing-jia SHEN</creatorcontrib><title>家蚕miR-0001和miR-0015体外下调BmFib-L基因的表达</title><title>浙江大学学报：B卷英文版</title><addtitle>Journal of Zhejiang University Science</addtitle><description>目的：探究新发现的两个miRNA对家蚕丝素轻链基因BmFib-L的调控作用。创新点：在家蚕后部丝腺中发现两个新的miRNA,并首次证明它们对家蚕丝素蛋白轻链基因BmFib-L有负调控作用。方法：本实验通过生物信息学分析,从家蚕后部丝腺miRNA高通量测序获得的新miRNA中,筛选出两个可能对家蚕丝素蛋白轻链基因BmFib-L有调控作用的miRNA,即bmo-miR-0001和bmo-miR-0015。设计茎环引物,采用反转录聚合酶链反应（RT-PCR）方法对家蚕5龄3 d头部、表皮、脂肪体、马氏管、精巢/卵巢、丝腺（前、中、后）、气管、中肠和血淋巴细胞等12个不同组织的bmo-miR-0001和bmo-miR-00015进行半定量表达分析。并采用双荧光报告基因检测系统进一步在细胞水平上验证bmo-miR-0001和bmo-miR-0015对BmFib-L表达的调控作用。结论：本实验中RT-PCR结果显示,这两个新miRNA在家蚕后部丝腺中表达量最高（图4）。双荧光报告基因检测结果显示,报告基因荧光素酶活性明显低于阳性对照组（图7）,转染bmo-miR-0001和bmo-miR-0015表达载体的细胞,报告基因和荧光素酶活性分别只及对照组60%和69%。t检验分析结果显示两个实验组与对照组之间差异都达到显著水平（P〈0.05）。由此可见,bmo-miR-0001和bmo-miR-0015在体外对BmFib-L的表达具有显著的抑制作用。</description><subject>BmFib-L</subject><subject>bmo-miR-0001</subject><subject>bmo-miR-0015</subject><subject>miRNA</subject><subject>功能验证</subject><subject>家蚕</subject><issn>1673-1581</issn><issn>1862-1783</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><recordid>eNpjYuA0tDAz0jU0tzBmAbLNzI11DU0tDDkYuIqLswwMTEwMzM04Gayfrtv2YtbU3MwgXQMDA8Onk3ogTEPTJ3snP10y7cmO7hcbmp1y3TKTdH2ezt_1dPaC57NaXixc8WLfPh4G1rTEnOJUXijNzaDk5hri7KGbnJGfl16YmZceX1CUmZtYVBlvZmZuaQx0j4ExUYoA-pdB_w</recordid><startdate>2016</startdate><enddate>2016</enddate><creator>Chen CHEN Yang-yang FAN Xin WANG Fei SONG Tao JIANG Ping QIAN Shun-ming TANG Xing-jia SHEN</creator><scope>2RA</scope><scope>92L</scope><scope>CQIGP</scope><scope>~WA</scope></search><sort><creationdate>2016</creationdate><title>家蚕miR-0001和miR-0015体外下调BmFib-L基因的表达</title><author>Chen CHEN Yang-yang FAN Xin WANG Fei SONG Tao JIANG Ping QIAN Shun-ming TANG Xing-jia SHEN</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-chongqing_primary_6679318603</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>BmFib-L</topic><topic>bmo-miR-0001</topic><topic>bmo-miR-0015</topic><topic>miRNA</topic><topic>功能验证</topic><topic>家蚕</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chen CHEN Yang-yang FAN Xin WANG Fei SONG Tao JIANG Ping QIAN Shun-ming TANG Xing-jia SHEN</creatorcontrib><collection>维普_期刊</collection><collection>中文科技期刊数据库-CALIS站点</collection><collection>维普中文期刊数据库</collection><collection>中文科技期刊数据库- 镜像站点</collection><jtitle>浙江大学学报：B卷英文版</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chen CHEN Yang-yang FAN Xin WANG Fei SONG Tao JIANG Ping QIAN Shun-ming TANG Xing-jia SHEN</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>家蚕miR-0001和miR-0015体外下调BmFib-L基因的表达</atitle><jtitle>浙江大学学报：B卷英文版</jtitle><addtitle>Journal of Zhejiang University Science</addtitle><date>2016</date><risdate>2016</risdate><volume>17</volume><issue>2</issue><spage>127</spage><epage>135</epage><pages>127-135</pages><issn>1673-1581</issn><eissn>1862-1783</eissn><notes>Chen CHEN, Yang-yang FAN, Xin WANG, Fei SONG, Tao JIANG, Ping QIAN, Shun-ming TANG, Xing-jia SHEN （Jiangsu Key Laboratory of Sericultural Biology and Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212003, China） （2Key Laboratory of Silkworm and Mulberry Genetic Improvement, Ministry of Agriculture, Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang 212018, China）</notes><notes>Based on bioinformatic analysis, we selected two novel micro RNAs（miRNAs）, bmo-miR-0001 and bmomiR-0015, from high-throughput sequencing of the Bombyx mori larval posterior silk gland（PSG）. Firstly, we examined the expression of bmo-miR-0001 and bmo-miR-0015 in 12 different tissues of the 5th instar Day-3 larvae of the silkworm. The results showed that the expression levels of both bmo-miR-0001 and bmo-miR-0015 were obviously higher in the PSG than in other tissues, implying there is a spatio-temporal condition for bmo-miR-0001 and bmo-miR-0015 to regulate the expression of Bm Fib-L. To test this hypothesis, we constructed pri-bmo-miR-0001 expressing the plasmid pc DNA3.0 [ie1-egfp-pri-bmo-miR-0001-SV40] and pri-bmo-miR-0015 expressing the plasmid pc DNA3.0 [ie1-egfp-pribmo-miR-0015-SV40]. Finally, the Bm N cells were harvested and luciferase activity was detected. The results showed that luciferase activity was reduced significantly（P〈0.05） in Bm N cells co-transfected by pc DNA3.0 [ie1-egfp-pri-bmomiR-0001-SV40] or pc DNA3.0 [ie1-egfp-pri-bmo-miR-0015-SV40] with p GL3.0 [A3-luc-Fib-L-3＇UTR-SV40], suggesting that both bmo-miR-0001 and bmo-miR-0015 can down-regulate the expression of Bm Fib-L in vitro.</notes><notes>Bombyx mori; MicroRNA; bmo-miR-0001; bmo-miR-0015; BmFib-L; Regulation of expression</notes><notes>33-1356/Q</notes><abstract>目的：探究新发现的两个miRNA对家蚕丝素轻链基因BmFib-L的调控作用。创新点：在家蚕后部丝腺中发现两个新的miRNA,并首次证明它们对家蚕丝素蛋白轻链基因BmFib-L有负调控作用。方法：本实验通过生物信息学分析,从家蚕后部丝腺miRNA高通量测序获得的新miRNA中,筛选出两个可能对家蚕丝素蛋白轻链基因BmFib-L有调控作用的miRNA,即bmo-miR-0001和bmo-miR-0015。设计茎环引物,采用反转录聚合酶链反应（RT-PCR）方法对家蚕5龄3 d头部、表皮、脂肪体、马氏管、精巢/卵巢、丝腺（前、中、后）、气管、中肠和血淋巴细胞等12个不同组织的bmo-miR-0001和bmo-miR-00015进行半定量表达分析。并采用双荧光报告基因检测系统进一步在细胞水平上验证bmo-miR-0001和bmo-miR-0015对BmFib-L表达的调控作用。结论：本实验中RT-PCR结果显示,这两个新miRNA在家蚕后部丝腺中表达量最高（图4）。双荧光报告基因检测结果显示,报告基因荧光素酶活性明显低于阳性对照组（图7）,转染bmo-miR-0001和bmo-miR-0015表达载体的细胞,报告基因和荧光素酶活性分别只及对照组60%和69%。t检验分析结果显示两个实验组与对照组之间差异都达到显著水平（P〈0.05）。由此可见,bmo-miR-0001和bmo-miR-0015在体外对BmFib-L的表达具有显著的抑制作用。</abstract></addata></record>
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subjects BmFib-L
bmo-miR-0001
bmo-miR-0015
miRNA
功能验证
家蚕
title 家蚕miR-0001和miR-0015体外下调BmFib-L基因的表达
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