盐生植物碱蓬Actin基因片段的克隆及序列分析

目的:克隆盐生植物碱蓬(Suaeda glauca)Actin基因片段,为研究其它基因在碱蓬的表达和调控提供内参基因.方法:根据已知植物Acfin基因的保守序列设计一对简并性引物,采用RT-PCR的方法扩增Actin基因片段,使用分子生物学软件进行序列分析.结果:获得一段大小为598bp的基因片段,编码198个氨基酸;该序列与其它Actin基因核苷酸序列的同源性均在80%以上,与氨基酸序列的同源性达93%以上.结论:克隆的基因为Actin基因片段,将其命名为SgACT,并登录在GenBank,登录号为EU429457.     

悬钩子属植物肌动蛋白基因片段的克隆与表达

目的:克隆悬钩子属植物肌动蛋白基因(actin),为研究该物种中其他基因的表达和调控提供内标基因.方法:利用一对特异性引物从黑莓、悬钩子杂种和树莓品种中克隆actin cDNA片段,对其进行序列分析和半定量RT-PCR表达分析.结果:从3个品种中均获得一条783 bp actin cDNA片段,编码260个氨基酸,3条actin片段与其他植物核苷酸序列的同源性都在82%以上,与其他植物氨基酸序列同源性也均在95%以上;树莓和悬钩子杂种品种的actin核苷酸和氨基酸序列同源性均达99%,系统进化分析也发现两者亲缘关系较近些;表达分析发现actin基因在各品种不同组织中均有一定的表达量.结论:首次克隆了悬钩子属植物肌动蛋白基因actin,将来自黑莓和树莓品种的序列登录在GenBank,登录号分别为HQ439556和HQ439557.     

木薯Actin基因片段的克隆及序列分析

克隆木薯Actin基因片段,为研究其他基因在木薯中的表达和调控提供内参基因.通过比较拟南芥、蓖麻和麻风树Actin基因cDNA同源区域,根据基因的保守序列设计一对简并性引物,采用RT-PCR的方法扩增Actin基因片段,使用分子生物学软件进行分析.结果显示,获得一段大小为698 bp的基因片段,编码233个氨基酸;该基因序列与其他Actin基因核苷酸序列的同源性均在85%以上,氨基酸序列的同源性达94%以上;系统进化分析表明,木薯Actin基因与大戟科植物橡胶、麻风树及蓖麻的亲缘关系最近,与毛果杨、陆地棉及木瓜等植物Actin基因具有较高的保守性.克隆的基因片段为木薯Actin基因片段,并命名为msACT.     

番杏Actin基因片段的克隆及生物信息学分析

本实验为研究番杏(Tetragonia tetragonioides)功能基因表达模式提供内参基因,根据登录在NCBI上植物的Actin基因的保守区域设计简并性引物,通过RT-PCR克隆获得番杏的Actin基因片段,将该片段连接于载体pGEM-T后进行测序,并将该基因序列通过生物信息学软件进行分析。结果显示,克隆所得基因片段大小为598 bp,编码198个氨基酸;该序列与登录在NCBI上的其他植物的Actin基因的核苷酸序列的同源性最大可达86%以上,编码蛋白的氨基酸序列同源性在88%以上。结果表明,本研究克隆所得的基因序列为番杏Actin基因片段。该基因命名为TtActin1,在Gen Bank的登录号为MH33308。     

蒙古冰草Actin基因片段的克隆及序列分析

旨在利用同源序列法分离蒙古冰草(Agropyron mongolicum Keng)Actin基因同源片段,为研究其他基因在蒙古冰草中的表达和调控提供内标参照.根据禾本科植物小麦Actin基因(AB181991)的保守序列设计2对引物A4和A5,采用RT-PCR扩增蒙古冰草的Actin基因片段,分别得到656 bp和848 bp的片段,使用DNAman和DNAuser等分子生物学软件进行序列分析,将2个片段的重复序列合并后获得一段长度为962 bp的基因片段,编码237个氨基酸,将克隆的Actin基因片段命名为MwACT.该序列与其它植物Actin基因核苷酸序列的同源性均在80%以上,其中与小麦、大麦的同源性达到94%;与氨基酸序列的同源性均在90%以上.     

厚藤Actin基因片段的克隆及序列分析

为研究功能基因在厚藤(Ipomoea pes-caprae L)中的表达和调控提供内参基因,本文根据Actin基因的保守区设计简并性引物,采用RT-PCR克隆厚藤的Actin基因片段,然后将获得的片段连接于克隆载体上进行测序,并运用分子生物学软件对该基因序列进行分析。结果表明,该基因片段大小为554bp,编码184个氨基酸;该序列与其他植物Actin基因的cDNA序列的同源性均在80%以上,与氨基酸序列的同源性在94%以上。由此得出,本研究克隆的基因序列为Actin基因片段,将其命名为IpActin1,并登录在GenBank,登录号为KU564627。     

滨梅2个肌动蛋白cDNA片段的克隆与表达

目的:克隆耐盐碱果树滨梅的肌动蛋白基因actin,为该物种优异性状基因的功能鉴定提供内参.方法:利用一对actin简并引物克隆滨梅actin cDNA片段,对其进行序列和表达分析.结果:得到2条743 bp的cDNA片段,两条核苷酸序列相似性为82％,命名为PmAct1和PmAct2并在GenBank登录(分别为JX855160和JX855161);序列比对发现2条actin片段氨基酸序列与其他植物同源性均在96％以上;根据不同果树Actin相似性构建进化树,表明2个滨梅actin明显分为两种类型,但均与蔷薇科果树亲缘关系较密切;半定量RT-PCR表达谱发现PmAct1可能为组成型表达类型,而PmAct2特异地在根和叶组织中表达量较高.结论:首次获得了2个滨梅actin基因,为该物种其他功能基因的挖掘和表达分析奠定了基础.     

滨海适生植物草海桐Actin基因片段的克隆及序列分析

[目的]本文为研究草海桐功能基因的表达模式提供可供参考的内参基因。[方法]根据Actin基因的保守区设计简并性引物,采用RT-PCR扩增草海桐Actin基因的片段。将获得的片段连接于T载体并进行测序,运用分子生物学软件对该序列进行分析。[结果]获得一段大小为554 bp的基因片段,编码184个氨基酸;该序列与其它Actin基因核苷酸序列的同源性均在82%以上,与氨基酸序列的同源性达96%以上。[结论]克隆的基因为Actin基因片段,将其命名为Ss Actin1,并登录在Gen Bank,登录号为KU564628。     

多浆旱生植物霸王Actin基因片段的克隆及序列分析

根据其他植物Actin基因的保守序列设计一对简并性引物,以霸王叶片总RNA为模板,采用RT-PCR的方法扩增出Actin基因片段并克隆到PUCm-T载体.阳性克隆经PCR鉴定后进行测序,序列分析结果表明:该片段长598bp,编码198个氨基酸;所得序列与GenBank中注册的Actin基因序列的同源性均在82%以上,与其他肌动蛋白的氨基酸序列的同源性达91%以上.     

拒盐型盐生植物小花碱茅肌动蛋白基因片段的克隆及序列分析

本研究根据其它植物Actin基因的保守序列设计一对简并性引物,以拒盐型盐生植物小花碱茅根部总RNA为模板,采用RT-PCR的方法扩增出Actin基因片段并克隆到PUCm-T载体,阳性克隆经PCR检测后进行测序,在GenBank中注册;序列分析结果表明:该片段长约600 bp,编码198个氨基酸;所得序列与GenBank中注册的其它植物Actin基因序列同源性均在84%以上,与其它肌动蛋白的氨基酸序列同源性达94%以上.     

Actin Gene Sequence from Euglena gracilis

ABSTRACT The full length coding sequence of the Euglena gracilis actin gene was determined by RT-PCR of Euglena gracilis mRNA. Conserved regions in the actin amino acid sequence were used as guides for the synthesis of degenerate primers. Sequence was obtained for 1.238 nucleotides, of which 1.131 were coding for 377 amino acids. Sequence comparisons showed a similarity with other actins of 56% to 80%. Even though most of the actin amino acid sequence was conserved, some regions showed high divergence, i.e. the DNase I-binding loop at the N-terminal region. The construction of a phylogenetic tree based on actin sequences from different organisms placed Euglena gracilis in a cluster with Trypanosoma brucei and Leishmania major.     

香蕉束顶病毒基因克隆和序列分析

对香蕉束顶病毒（ＢＢＴＶ）中国分离株ＤＮＡ组份Ｉ（ＤＮＡ－１）、外壳蛋白（ＣＰ）和运转蛋白（ＭＰ）基因进行了克隆和序列分析。ＢＢＴＶＤＮＡ－１含有１１０３个核苷酸,与南太平洋和亚洲分离株分别有８７％－８８％ ９６．９－９８％的核苷酸序列同源性。由ＤＮＡ－１编码的复制酶含有１８６个在酸残基。与南太平洋和亚洲分离株分别有８４．４％－９５．８％和９７．６％、９８．０％的氨基酸序列同源性。外壳蛋白基因由５     

水花生Actin基因片段的克隆及序列分析

肌动蛋白基因编码生物体内一种很重要的组成型表达蛋白质,常被看作看家基因。为分离水花生肌动蛋白基因,根据GenBank上一藜科植物的———Actin基因的EST序列,分别设计引物,提取水花生根的总RNA,对上述的基因片断进行RT—PCR,扩增后测序得到230bp长的核苷酸序列。经同源性比较,这条序列与上述藜科植物Actin的EST序列基本一致,表明获得了水花生肌动蛋白序列。     

Chemical composition and antioxidant activities of Broussonetia papyrifera fruits

Fruits of Broussonetia papyrifera from South China were analyzed for their total chemical composition, and antioxidant activities in ethanol and aqueous extracts. In the fruit of this plant, the crude protein, crude fat and carbohydrates was 7.08%, 3.72% and 64.73% of dry weight, respectively. The crude protein, crude fat and carbohydrates were 15.71%, 20.51% and 36.09% of dry weight, respectively. Fatty acid and amino acid composition of the fruit were analyzed. Unsaturated fatty acid concentration was 70.6% of the total fatty acids. The percentage of the essential amino acids (EAAs) was 40.60% of the total amino acids. Furthermore, B. papyrifera fruit are rich in many mineral elements and vitamins. Total phenolic content was assessed using the Folin-Ciocalteau assay, whereas antioxidant activities were assessed by measuring the ability of the two extracts to scavenge DPPH radicals, inhibit peroxidation, and chelate ferric ions. Their reducing power was also assessed. Results indicated that the aqueous extract of B. papyrifera was a more potent reducing agent and radical-scavenger than the ethanol extract. GC-MS analysis of the ethanol extract showed the presence of some acid-containing compounds. The changes in total phenolic content and antioxidant capacity in B. papyrifera from four different regions grown under normal conditions were assessed. The antioxidant activity of different extracts was positively associated with their total phenolic content. These results suggest that the fruit of B. papyrifera could be used in dietary supplement preparations, or as a food additive, for nutritional gain, or to prevent oxidation in food products.     

水稻条叶枯病毒(RStV)基因组组分4的克隆与序列分析

利用RTPCR技术合成并扩增了水稻条叶枯病毒(RStV)中国云南分离物基因组组分4的全长cDNA,将PCR产物克隆在载体pCRII上,并进行全序列测定,所得核苷酸序列及推测的氨基酸序列与日本分离物T进行比较。结果表明,在核苷酸水平,两分离物的vORF、vcORF及基因间非编码区序列的同源性分别为94.9%、94.1%、86.1%,5’端非编码区序列相同,而3’非编码区同源性为96.1%,仅有两个核苷酸不同;在氨基酸水平,vORF及vcORF编码蛋白的同源性分别为99.4%和98.3%。可见,编码区的大小及其氨基酸序列和两末端序列都是很保守的。因此,中国云南分离物Y与日本分离物T可能有很近的亲缘关系。     

The budding yeast U5 snRNP Prp8 is a highly conserved protein which links RNA splicing with cell cycle progression.

The dbf3 mutation was originally obtained in a screen for DNA synthesis mutants with a cell cycle phenotype in the budding yeast Saccharomyces cerevisiae. We have now isolated the DBF3 gene and found it to be an essential gene with an ORF of 7239 nucleotides, potentially encoding a large protein of 268 kDa. We also obtained an allele-specific high copy number suppressor of the dbf3-1 allele, encoded by the known SSB1 gene, a member of the Hsp70 family of heat shock proteins. The sequence of the Dbf3 protein is 58% identical over 2300 amino acid residues to a predicted protein from Caenorhabditis elegans. Furthermore, partial sequences with 61% amino acid sequence identity were deduced from two files of human cDNA in the EST nucleotide database so that Dbf3 is a highly conserved protein. The nucleotide sequence of DBF3 turned out to be identical to the yeast gene PRP8, which encodes a U5 snRNP required for pre-mRNA splicing. This surprising result led us to further characterise the phenotype of dbf3 which confirmed its role in the cell cycle and showed it to function early, around the time of S phase. This data suggests a hitherto unexpected link between pre-mRNA splicing and the cell cycle.     

猪骨骼肌快肌肌钙蛋白C2基因的cDNA克隆与表达分析

从人骨骼肌快肌肌钙蛋白C2（TNNC2）基因出发,在dbEST数据库中进行同源性搜索,找到一个有较高同源性且在猪背最长肌中表达EST（BM083186）。通过电子克隆和进一步RT-PCR实验验证,获得猪TNNC2基因全长cDNA序列,其全长843bp,开放阅读框为201～683bp,编码有160个氨基酸。同源性分析结果表明,与人、鼠的骨骼肌快肌肌钙蛋白C2基因cDNA编码区（CDS）同源性分别为93.6％、90.5％,蛋白序列同源性均为97.5%。多种组织的半定量RT-PCR研究表明,该基因在骨骼肌中表达,并且在杜洛克猪背最长肌中的表达比兰塘猪高。     

Two branches of the lupeol synthase gene in the molecular evolution of plant oxidosqualene cyclases.

Two new triterpene synthase cDNAs, named as OEW and TRW, were cloned from olive leaves (Olea europaea) and from dandelion roots (Taraxacum officinale), respectively, by the PCR method with primers designed from the conserved sequences found in the known oxidosqualene cyclases. Their ORFs consisted of 2274 bp nucleotides and coded for 758 amino acid long polypeptides. They shared high sequence identity (78%) to each other, while they showed only about 60% identities to the known triterpene synthases LUPI (lupeol synthase clone from Arabidopsis thaliana) and PNY (beta-amyrin synthase clone from Panax ginseng) at amino acid level. To determine the enzyme functions of the translates, they were expressed in an ERG7 deficient yeast mutant. Accumulation of lupeol in the cells of yeast transformants proved both of these clones code for lupeol synthase proteins. An EST (expression sequence tag) clone isolated from Medicago truncatula roots as a homologue of cycloartenol synthase gene, exhibits high sequence identity (75-77%) to these two lupeol synthase cDNAs, suggesting it to be another lupeol synthase clone. Comparatively low identity (approximately 57%) of LUP1 from Arabidopsis thaliana to either one of these clones leaves LUP1 as a distinct clone among lupeol synthases. From these sequence comparisons, now we propose that two branches of lupeol synthase gene have been generated in higher plants during the course of evolution.     

杨树粒肩天牛的生物学特性

杨树粒肩天牛在福建省2a发生1代;老熟幼虫在第2年4月下旬至5月初开始化蛹,成虫出现期始于5月底6月初,6月中下旬到7月上旬为盛发期,9月上旬只见个别成虫;产卵期始于6月中旬,6月下旬至7月中旬为产卵高峰期;幼虫孵化期为6月下旬到8月中旬,7月上中旬为为盛发期;幼虫不越冬。杨树粒肩天牛成虫为补充营养所取食的树种较集中,主要是构树与桑树。杨树粒肩天牛幼虫对杨树危害严重,但成虫却不喜欢取食杨树,用杨树饲养的粒肩天牛成虫寿命很短,仅3-19d,不产卵;而以桑树为补充营养的雌成虫的平均寿命为55d、雄26.5d;以构树为补充营养的雌成虫的平均寿命为78d、雄45.5d,补充桑树与构树的粒肩天牛成虫均可正常产卵。雌雄成虫一生可多次交尾。雌虫有多次产卵现象,单雌每天产卵数量1-12粒不等。