草鱼肠系膜脂肪组织高质量总RNA提取方法的研究     

姜鹏  骆丽婷  李胜杰  樊佳佳  马冬梅 《水产科学》2021,(2):166-171

基于传统的酸性酚—异硫氰酸胍—氯仿一步提取法,比较分析多种优化操作步骤,摸索出一种高质量提取体质量为80~150 g草鱼肠系膜脂肪组织总RNA的改良方法。试验结果显示,相较于肝脏、脾脏、肠道等脏器组织,草鱼肠系膜脂肪组织RNA丰度低,且极易在样品前处理阶段出现顽固性降解问题。探索发现,将取样量增至约30 mg,可提升RNA产量以满足常规试验需求。针对降解难题,改良常规的样品前处理技术流程,采用鲜样液氮速冻,冻样直接放入TRIzol试剂中裂解,并即刻进行长时间机械匀浆,时长约3 min等核心操作步骤,可显著降低脂肪组织样品RNA的降解。Agilent生物分析仪检测结果显示,改良方法提取的草鱼肠系膜脂肪组织RNA完整度高,关键RIN值为8.7~9.0。研究推测,长时间机械匀浆所形成的持续剪切冲击力或许有助于TRIzol试剂中的异硫氰酸胍等成分突破油滴阻碍而有效抑制内源性RNA酶。本方法提取的草鱼脂肪组织总RNA质量可满足高通量转录组测序要求。  相似文献   

Transcriptome Analysis of Myzus persicae to UV-B Stress     

Chang-Li Yang  Jian-Yu Meng  Meng-Shuang Yao  Chang-Yu Zhang 《Journal of insect science (Online)》2021,21(3)

  相似文献   

Identification,characterization and full-length sequence analysis of a novel endornavirus in common sunflower (Helianthus annuus L.)     

Wen-wen LIU  Min XIN  Meng-ji CAO  Meng QIN  Hui LIU  Shou-qi ZHAO  Xi-feng WANG 《农业科学学报》2018,17(10):2281-2291

To identify the possible quarantine viruses in seven common sunflower varieties imported from the United States of America and the Netherlands, we tested total RNAs extracted from the leaf tissues using next-generation sequencing of small RNAs. After analysis of small RNA sequencing data, no any quarantine virus was found, but a double-stranded RNA(dsRNA) molecule showing typical genomic features of endornavirus was detected in two varieties, X3939 and SH1108. Full-length sequence and phylogenetic analysis showed that it is a novel endornavirus, temporarily named as Helianthus annuus alphaendornavirus(HaEV). Its full genome corresponds to a 14 662-bp dsRNA segment, including a 21-nt 5′ untranslated region(UTR), 3' UTR ending with the unique sequence CCCCCCCC and lacking a poly(A) tail. An open reading frame(ORF) that encodes a deduced 4 867 amino acids(aa) polyprotein with three domains: RdRP, Hel and UGT(UDP-glycosyltransferase). HaEV mainly distributed in the cytoplasm but less in the nucleus of leaf cells by fluorescence in situ hybridization(FISH) experiment. This virus has a high seed infection rate in the five varieties, X3907, X3939, A231, SH1108 and SR1320. To our knowledge, this is the first report about the virus of the family Endornaviridae in the common sunflower.  相似文献   

以塘坝为中心建设立体型综合开发小区     

于明 《中国水土保持》1994,(1):49-50

以塘坝为中心建设立体型综合开发小区于明（黑龙江省青冈县水土保持办公室，１５１６０６）塘坝是我国水利水保建设的重要工程措施之一。它不仅可防洪蓄水，防治水土流失，同时对解决农田抗旱灌溉、养鱼等综合开发利用具有重要作用。搞好以塘坝建设为中心的小流域综合开发...  相似文献   

T7 RNA多聚酶基因的克隆、序列测定及其在E.coli中的表达     

余兴龙  涂长春  徐兴然  张茂林  张青婵  李作生 《中国兽医学报》2003,23(5):452-454

从BL21(DE3)E．coli菌株中以PCR的方法扩增得到了与T7RNA多聚酶(T7RNApolymerase，T7pol)基因大小一致的DNA片断。将PCR产物纯化后直接克隆到pGEM—T载体中，经酶切鉴定和DNA序列分析表明克隆得到了正确的T7pol基因。将T7pol基因亚克隆入pET-28b( )中，构建得到原核表达质粒pET28T7。该质粒的BL21(DE3)pLysS转化菌在IPTG的诱导下可表达约98800的蛋白，这与T7pol的相对分子质量一致。将该质粒转化DH5α、JMl09、HBl01、BL21(DE3)和BL21(DE3)pLysS等5种不同的宿主菌，仅有转化T7pol酶活性受到抑制的宿主菌BL21(DE3)pLysS才能得到转化子，而其余4种T7T7pol酶活性不受抑制的E．coli宿主菌不能得到转化子。pET28T7原核表达质粒这种仅能在T7pol酶活性受到抑制的宿主菌中才能存活的现象说明本试验所克隆的T7pol基因能正确表达出具有RNA转录酶活性的蛋白。  相似文献   

The RNA interference     

KONG Xiang-ping  Lixing W. Reneker 《园艺学报》2004,20(5):900-903

The phenomenon of RNA interference (RNAi) is highly conserved mechanism in the organism evolution. As a immune system, RNAi is a ubiquitous mechanism against invading microorganism in plant and animal cells. Recently, it has been found that RNAi is the process by which double-strand RNA(dsRNA) directs sequence-specific degradation of messenger RNA and the mediations of sequence specific messenger RNA degradation are 21-and 23-nucleotide small interfering RNAs that generate by ribonuclease from endogenous longer dsRNA or by transfectious technics from heterologous dsRNA. Over the past few years, the way in which cells respond to dsRNA by silencing homologous genes has revealed a new regulating paradigm in biology.  相似文献   

The Application of GIS in Small Watershed Classification in Loess Plateau     

Zhu Jinzhao  Wu Bin  Bi Huaxing  Zhou Changqing College of Soil  Water Conservation  Beijing Forestry UniversityBeijing  P.R.China 《中国林学(英文版)》1994,(2)

ＴｈｅＡｐｐｌｉｃａｔｉｏｎｏｆＧＩｓｉｎＳｍａｌｌＷａｔｅｒｓｈｅｄＣｌａｓｓｉｆｉｃａｔｉｏｎｉｎＬｏｅｓｓＰｌａｔｅａｕＺｈｕＪｉｎｚｈａｏ，ＷｕＢｉｎ，ＢｉＨｕａｘｉｎｇ，ＺｈｏｕＣｈａｎｇｑｉｎｇＣｏｌｌｅｇｅｏｆＳｏｉｌａｎｄＷａｔｅｒＣ...  相似文献   

对小水电、沼气池、省柴灶三个能源工程项目的综合评价   总被引：1，自引：0，他引：1  

曾晶 《贵州农业科学》2005,33(4):65-67

农村能源工程项目的建设不仅具有项目本身的经济效益，而且有更广泛的能源、间接的经济和生态效益。农村能源工程项目的建设除了受资金约束外，还有技术管理等方面的限制，甚至人们的习俗和心理状态都可能是能源工程项目的制约因素。因此，对农村能源工程项目的多种效益和多种制约的综合分析和评价，将使我们能更全面地考虑项目的综合效果，使有限的资金、人力、物力投入到综合效果最好的项目上去。  相似文献   

新疆水土保持情况调查     

郭廷辅  佟伟力 《中国水土保持》1994,(12):5-8

新疆水土保持情况调查郭廷辅，佟伟力（水利部水土保持司，北京１００７６１）我们于１９９４年８月１１日至２６日对新疆伊犁、和田、喀什、吐鲁番、昌吉、乌鲁木齐等６个地区（州、市）的１１个县（市）、两个流域的水土流失情况和开展治理工作做了典型调查。同时，也对...  相似文献   

杂交条纹鲈苗种小水泥池培育技术试验     

田功太段登选  曹振杰马荣棣  陈述江孙栋 陈金萍 《齐鲁渔业》2004,21(7):1-3

用地上露天小水泥池8个．进行杂交条纹鲈苗种培育试验．放养313龄水花30000尾．通过天然饵料、藻粉投喂以及适当水质管理措施．经32天培育，共培育3．5～4．6cm体长的鱼种4336尾．最高成活率为19．1％，最低为6％，平均为14．5％。  相似文献