Chinese tomato yellow leaf curl virus--a new species of geminivirus

ＧｅｍｉｎｉｖｉｒｕｓｅｓａｒｅａｇｒｏｕｐｏｆｐｌａｎｔｖｉｒｕｓｅｓｃｈａｒａｃｔｅｒｉｚｅｄｂｙｔｈｅｉｒｃｉｒｃｕｌａｒｓｉｎｇｌｅｓｔｒａｎｄｅｄＤＮＡ(ｓｓＤＮＡ)ｇｅｎｏｍｅａｎｄａｕｎｉｑｕｅｇｅｍｉｎａｔｅｐａｒｔｉｃｌｅｍｏｒｐｈｏｌｏｇｙ[1].Ｇｅｍｉｎｉｖｉｒｕｓｅｓａｒｅｄｉｖｉｄｅｄｉｎｔｏｔｈｒｅｅｓｕｂｇｒｏｕｐｓｏｎｔｈｅｂａｓｉｓｏｆｇｅｎｏｍｅｏｒｇａｎｉｚａｔｉｏｎａｎｄｉｎｓｅｃｔｖｅｃｔｏｒ:ＡｌｌｓｕｂｇｒｏｕｐＩｇｅｍｉｎｉｖｉｒｕｓｅｓａｒｅｌｅａｆ…     

DNA分子标记在柑桔中的应用

ＤＮＡ分子标记是最为理想的遗传标记 ,依其多态性检出所用的分子生物学技术 ,大致可分为Ｓｏｕｔｈｅｍ杂交技术为核心的分子标记和ＰＣＲ技术为核心的分子标记。前者的代表性技术有ＲＥＬＰ(ｒｅｓｔｒｉｃｔｉｏｎｆｒａｇｍｅｎｔｌｅｎｇｔｈｐｏｌｙｍｏｒｐｈｉｓｍ)和ＤＮＡ指纹技术 (ＤＮＡｆｉｎｇｅｒｐｒｉｎｔｉｎｇｔｅｃｈｎｉｑｕｅｓ)。后者的代表性技术有ＲＡＰＤ(ｒａｎｄｏｍａｍｐｌｉｆｉｅｄｐｏｌｙｍｏｒｐｈｉｃＤＮＡ)、ＳＣＡＲ(ｓｅｑｕｅｎｃｅｃｈａｒａｃ ｔｅｒｉｓｔｉｃａｍｐｌｉｆｉｅｄｒｅｇｉｏｎ)…     

莴苣叶绿体psbA基因序列

1　Ｓｏｕｒｃｅ　ＴｈｅｓｅｑｕｅｎｃｅｗａｓｄｅｔｅｒｍｉｎｅｄｆｒｏｍａＰＣＲｐｒｏｄｕｃｔｏｆｃｈｌｏｒｏｐｌａｓｔｇｅｎｏｍｉｃＤＮＡｏｆＬａｃｔｕｃａｓａｔｉｖａ (ｌｅｔｔｕｃｅ) .2　Ｎａｍｅａｎｄｄｅｓｃｒｉｐｔｉｏｎ　Ｔｈｅｃｏｄｏｎｓｅｑｕ     

油菜叶绿体基因ndhB的序列

1　Ｓｏｕｒｃｅ　ＴｈｅｓｅｑｕｅｎｃｅｗａｓｄｅｔｅｒｍｉｎｅｄｆｒｏｍａＰＣＲ ｐｒｏｄｕｃｔ,ｗｈｉｃｈｗａｓｌｉｇａｔｅｄｔｏｐＢｌｕｅｓｃｒｉｐｔＳＫ( )ｖｅｃｔｅｒ (ｎａｍｅｄａｓｐＳＮ) ,ｆｒｏｍｔｈｅｃｈｌｏｒｏｐｌａｓｔｇｅｎｏｍｉｃＤＮＡｏｆＢｒａｓｓｉｃａｎａｐｕｓ(Ｏｉｌｓｅｅｄｒａｐｅ)ｃｖ.Ｈ1 65.2　Ｎａｍｅａｎｄｄｅｓｃｒｉｐｔｉｏｎ　Ｂｒａｓｓｉｃａｎａｐｕｓｃｈｌｏｒｏ ｐｌａｓｔｇｅｎｅｎｄｈＢｉｓ 2 4 67ｂｐｌｏｎｇｗｉｔｈｔｗｏｅｘｏｎｓｔｈａｔｅｎｃｏｄｅ 51 0ａ…     

诸葛菜基因EPSPS的cDNA核苷酸序列

1　Ｓｏｕｒｃｅ　Ｔｈｅｓｅｑｕｅｎｃｅｗａｓｄｅｔｅｒｍｉｎｅｄｕｓｉｎｇｔｈｅ3′ＲＡＣＥ(ｒａｐｉｄａｍｐｌｉｆｉｃａｔｉｏｎｃＤＮＡｅｎｄｓ)ＲＴ ＰＣＲａｎｄ 5′ＲＡＣＥＲＴ ＰＣＲ ｐｒｏｄｕｃｔ,ｗｈｉｃｈｗａｓｌｉｇａｔｅｄｔｏｔｈｅ ｐＭＤ1 8 Ｔ     

Influence of loop sequence on relative stability of bimolecular triplex DNA

ＨｏｍｏｐｙｒｉｍｉｄｉｎｅｏｌｉｇｏｎｕｃｌｅｏｔｉｄｅｓｃａｎｂｉｎｄｔｏｔｈｅｍａｊｏｒｇｒｏｏｖｅｏｆｄｕｐｌｅｘＤＮＡａｔｈｏｍｏｐｕｒｉｎｅｈｏｍｏｐｙｒｉｍｉｄｉｎｅｓｅｑｕｅｎｃｅｓ,ｆｏｒｍｉｎｇａｌｏｃａｌｔｒｉｐｌｅｈｅｌｉｘ[1—3].ＴｈｅｒｅａｒｅｓｏｍｅｋｉｎｄｓｏｆｔｒｉｐｌｅｘＤＮＡ,ｕｎｉｍｏｌｅｃｕｌａｒｏｒｂｉｍｏｌｅｃｕｌａｒｏｒｔｅｒｍｏｌｅｃｕｌａｒｔｒｉｐｌｅｘｗｈｉｃｈｃｏｎｓｉｓｔｓｏｆｏｎｅｏｒｔｗｏｏｒｔｈｒｅｅｓｔｒａｎｄｓｏｆＤＮＡ,ｒｅｓｐｅ…     

小麦紫黄质脱环氧化酶基因WVDE序列

1　Ｓｏｕｒｃｅ　ＴｈｅｇｅｎｅｗａｓｉｓｏｌａｔｅｄｆｒｏｍＴｒｉｔｉｃｕｍａｅｓ ｔｉｖｕｍ (ｓｔｒａｉｎ:Ｈａｎｆｅｎｇ 970 3)ｌｅａｆｃＤＮＡｂｙＰＣＲ .2　Ｎａｍｅａｎｄｄｅｓｃｒｉｐｔｉｏｎ　ＴｈｅＴｒｉｔｉｃｕｍａｅｓｔｉｖｕｍｖｉｏｌａｘａｎｔｈｉｎｄｅ ｅｐｏｘｉｄａｓｅｇｅｎｅ (ｎａｍｅｄＷＶＤＥ)ｉｓ1 746ｂｐｌｏｎｇａｎｄｅｎｃｏｄｅｓ 45 4ａｍｉｎｏａｃｉｄｓｏｆｖｉｏｌａｘ ａｎｔｈｉｎｄｅ ｅｐｏｘｉｄａｓｅ .ＴｈｅｎｕｃｌｅｏｔｉｄｅｓｅｑｕｅｎｃｅｏｆＷＶＤＥｓｈｏｗｓ 83…     

Identification and characterization of scaffold-associated region (SAR) of rRNA gene of silkworm Attacus ricini

Ｓｃａｆｆｏｌｄａｓｓｏｃｉａｔｅｄｒｅｇｉｏｎ(ＳＡＲ)ａｎｄｍａｔｒｉｘａｔｔａｃｈｍｅｎｔｒｅｇｉｏｎ(ＭＡＲ)ａｒｅｋｎｏｗｎａｓｃｉｓａｃｔｉｎｇＤＮＡｅｌｅｍｅｎｔｓ,ｗｈｉｃｈｓｐｅｃｉｆｉｃａｌｌｙｂｉｎｄｔｏｎｕｃｌｅａｒｓｃａｆｆｏｌｄｏｒｎｕｃｌｅａｒｍａｔｒｉｘ.ＳＡＲ/ＭＡＲａｒｅｕｓｕａｌｌｙ300—1000ｂｐｌｏｎｇ,ＡＴｒｉｃｈ,ａｎｄｃｏｎｔａｉｎｉｎｇｔｏｐｏｉｓｏｍｅｒａｓｅＩＩｃｏｎｓｅｎｓｕｓｓｅｑｕｅｎｃｅａｎｄｏｔｈｅｒＡＴｒｉｃｈｓｅｑｕｅｎｃｅｍｏｔｉｆｓ[1].…     

DIFFERENT RESULTS BY DIFFERENT COMMERCIAL TAQ DNA POLYMERASE IN RAPD

ＲＡＰＤ (ＲａｎｄｏｍＡｍｐｌｉｆｉｅｄＰｏｌｙｍｏｒｐｈｉｃＤＮＡ )ｔｅｃｈｎｉｑｕｅｈａｓｂｅｅｎｗｉｄｅｌｙｕｓｅｄｉｎａｎｉｍａｌ,ｐｌａｎｔ,ｈｕｍａｎａｎｄｍｉｃｒｏｏｒｇａｎｉｓｍｒｅｓｅａｒｃｈｓｉｎｃｅｉｔｗａｓｆｉｒｓｔｅｓｔａｂｌｉｓｈｅｄｂｙＷｉｌｌｉａｍｓｉｎ 1 990 [1-3 ] .Ｂｕｔ,ｂｅｃａｕｓｅｏｆｌｏｗａｎｎｅａｌｅｄｔｅｍｐｅｒａｔｕｒｅａｎｄｓｈｏｒｔ1 0 -ｎｔｐｒｉｍｅｒｓ,ｔｈｅｒｅｓｏｌｕｔｉｏｎａｎｄｒｅｐｅｔｉｔｉｏｎｉｓｌｏｗｉｎＲＡＰＤ .Ｔｈｅｓｔａｂｉ…     

核基质结合区(MARs)与植物转基因的表达

基质结合区 (ｍａｔｒｉｘａｓｓｏｃｉａｔｉｏｎｒｅｇｉｏｎｓｏｒｍａｔｒｉｘａｔｔａｃｈｍｅｎｔｒｅ ｇｉｏｎｓ,ＭＡＲｓ)亦称核骨架结合区 (ｓｃａｆｆｏｌｄａｓｓｏｃｉａｔｉｏｎｒｅｇｉｏｎｓｏｒｓｃａｆｆｏｌｄａｔｔａｃｈｍｅｎｔｒｅｇｉｏｎｓ,ＳＡＲｓ) ,是真核基因组中能与核基质特异紧结合的ＤＮＡ序列。核基质与生物体内许多重要生命活动相联系 ,而结合的ＭＡＲｓ序列 ,参与ＤＮＡ复制 ,基因转录 ,基因表达调控[1 ] 和基因边界定位[2 ] 等过程。ＭＡＲｓ通过将染色质锚定在核基质上而使染色质形成拓扑学限制性Ｄ…     

人尿道(阴茎)鳞癌上皮细胞(HUS-98)cDNA文库的构建及鉴定

为了进一步分离人尿道(阴茎)鳞癌组织特异性表达基因和鳞癌特异性相关基因,采用SMART技术,构建了人尿道 (阴茎)鳞癌上皮细胞cDNA文库,从人尿道(阴茎)鳞癌上皮细胞中分离总RNA并纯化mRNA,利用经修饰的oligo(dT)引物 合成cDNA第一链,利用SMART核苷酸作为cDNA第一链在mRNA5′端延伸出去的模板,采用LD-PCR合成双链cDNA,双链 cDNA经酶切和过柱分级分离后,克隆入λTriplEx2载体后经体外包装而成cDNA文库。结果表明原始人尿道(阴茎)鳞癌上 皮cDNA文库获得1.57×107个重组子,重组率达到98%。文库扩增后,滴度达到4.0×109pfu/ml,插入cDNA平均长度为2.5kb。 构建的人尿道(阴茎)鳞癌上皮cDNA文库具有良好的质量,该cDNA文库为进一步筛选鳞癌抑癌基因及鳞癌特异性表达基因 奠定了基础。     

cDNA cloning of a mannose-binding lectin-associated serine protease (MASP) gene from hagfish (Eptatretus burgeri)

Hagfish, agnathan cyclostome, is the most primitive extant vertebrate and its complement (C) system seems to be a primordial system in comparison with a well-developed C system in gnathostome vertebrates. From a phylogenic perspective of defense mechanisms, we have isolated complement C3 from the serum of hagfish (Eptatretus burgeri). In this study, we first attempted to identify a hagfish Bf or C2 as a C3 convertase by RT-PCR using degenerative primers designed on the basis of the conserved amino acid stretches among the several kinds of serine proteases. Contrary to our expectation, homology search of cloned RT-PCR product suggested that there was a partial cDNA encoding the homologue of neither Bf nor C2 but a mannose-binding lectin-associated serine protease (MASP). Analyses of a full-length cDNA clone isolated from a hagfish liver cDNA library by using the partial cDNA as a probe indicated that this cDNA encoded hagfish MASP 1. This evidence strongly suggests that the hagfish defends itself against pathogens at least by the complement system composed of lectin pathway.     

Bovine corneal protein 54K (BCP54) is a homologue of the tumor-associated (class 3) rat aldehyde dehydrogenase (RATALD)

Amino acid (aa) sequence data from Staphylococcus areas V8 protease-digested bovine corneal 54-kDa protein (BCP54) fragments were utilized to derive mixed oligodeoxyribonucleotide (oligo) primers complementary to the reverse translation products of these sequences. These degenerate oligo primers were used to prime the amplification of BCP54 sequence from bovine corneal epithelial cell cDNA. The cDNA probe generated by this mixed oligo-primed amplification of cDNA was cloned and dideoxy-sequenced. A search of the GenBank database (version 63.0) revealed extensive sequence similarity to the cDNA encoding tumor-associated rat liver (class 3) aldehyde dehydrogenase (RATALD). Nucleotide (nt) and aa sequence alignment of the BCP54 translation product reveals it is 78% and 84% homologous with RATALD at the nt and aa levels, respectively. Conservation of aa sequence elements common to the aldehyde dehydrogenase family thought to be of structural/functional significance is further substantiated by this analysis. Included in the discussion is the likelihood that gene sharing (genes encoding metabolic enzymes and other stable proteins) may extend to the cornea.     

家蝇cDNA文库的构建

自Ｂｏｍａｎ小组 (Ｓｔｅｉｎｅｒ ,1981)从惜古比天蚕(Ｈｙａｌｏｐｈｏｒａｃｅｃｒｏｐｉａ)中分离、纯化出第一种抗菌肽天蚕素以来 ,人们已在昆虫和其他无脊椎动物中发现了 170多种抗菌肽 (Ｌｏｗｅｎｂｅｒｇｅｒｅｔａｌ ,1999)。目前 ,人们不仅搞清楚了多数抗菌肽的氨基酸序列、结构和功能特点 ,而且还对一些抗菌肽的基因进行了克隆 (陈留存和王金星 ,1999)。我们以家蝇为材料 ,分离、纯化出了抗菌肽 ,在此基础上 ,构建了经过诱导的家蝇ｃＤＮＡ文库 ,以克隆其基因。1　材料和方法1 1　实验材料家蝇 (Ｍｕｓｃａｄｏｍｅｓｔｉｃａ)…     

灰飞虱高带毒（RSV）群体酵母双杂交cDNA文库的构建

为了研究灰飞虱Laodelphax striatellus Fallén与水稻条纹病毒（rice stripe virus, RSV）互作机制, 本研究构建了灰飞虱高带毒群体酵母双杂交cDNA文库。以实验室筛选的灰飞虱高带毒群体为材料, 分离纯化mRNA, 反转录合成双链cDNA, 并连接三框型接头, 层析柱分级纯化。采用同源重组反应制备三框型cDNA入门文库, 再通过同源重组将入门文库转移到Gateway兼容载体pGADT7-DEST上, 构建获得酵母双杂交cDNA文库。检测结果表明： 文库库容量为3.68×10⁷ cfu, 扩增文库滴度为2.62×10¹⁰ cfu/mL; 文库重组率大于95%, cDNA插入片段平均长度>1 kb, 达到了标准cDNA文库的要求。灰飞虱高带毒群体酵母双杂交cDNA文库的构建为开展昆虫介体与水稻条纹病毒互作机制的研究奠定了基础。     

Cloning,sequencing and phylogenetic/phenetic classification of an estrogen receptor alpha (alpha) subtype of sheepshead minnow (Cyprinodon variegatus)

The estrogen receptor (ER) is a key component of the reproductive system of both teleosts and tetrapods. In this study, the sequence and evolutionary relationship of sheepshead minnow (Cyprinodon variegatus) ER were examined. Total RNA from livers of adult laboratory-reared gravid female C. variegatus was reverse-transcribed to prepare cDNA. Nested pairs of gene-specific degenerative primers derived from conserved amino acid sequences of the ER DNA binding domain were used to amplify an internal fragment of the ER cDNA of C. variegatus using polymerase chain reaction (PCR) followed by rapid amplification of cDNA ends (RACE). The amplified cDNA products were inserted into pGEM T-Easy Vector for cloning and sequencing. The cloned ER cDNA segments gave a 524-amino-acid ER sequence, which represents approximately 80% of the sequence. The use of PHYLIP for phylogenetic analysis with the maximum parsimony method and for phenetic analysis with the neighbor-joining method, along with bootstrap resampling, using 24 known sequences of alpha and beta ER subtypes (both teleosts and tetrapods) indicated that the ER cDNA sequence of C. variegatus has strong homology to the alpha-subtype (ER alpha) of other teleostean fish, especially the closely related killifish species, Japanese medaka (Oryzias latipes). Because ER alpha was the only subtype found, it appears that the alpha-subtype is predominant in C. variegatus liver.     

山羊发情相关输卵管糖蛋白(gEGP)基因的克隆

哺乳动物的输卵管精心设计了一个独特的液体环境 ,使得雌、雄性生殖细胞的运输及最终成熟、受精和早期胚胎发育能够顺利进行 .其中有一种源于分泌细胞的蛋白—发情相关的输卵管糖蛋白 (ｅｓｔｒｕｓ ａｓｓｏｃｉａｔｅｄｏｖｉｄｕｃｔｕａｌｇｌｙｃｏｐｒｏｔｅｉｎ ,ＥＧＰ) ,自从Ｓｕｔｔｏｎ等于 1984年首次在绵羊中发现的[1] ,在其他的物种中 ,如 ,狒狒、人、小鼠、猕猴、牛、羊、猪、仓鼠等 ,也都有发现[2 ,3 ,4] .虽然已有实验数据证明 ,当卵子经过输卵管时 ,它们可与卵子的透明带 (ｚｏｎａｐｅｌｌｕｃｉｄａ ,ＺＰ)和 或卵黄周…     

Human complex II (succinate-ubiquinone oxidoreductase): cDNA cloning of iron sulfur (Ip) subunit of liver mitochondria

Complex II (succinate-ubiquinone oxidoreductase) is an important enzyme complex of both the tricarboxylic acid cycle and of the aerobic respiratory chains of mitochondria in eukaryotic cell and prokaryotic organisms. In this study, the amino acid sequence of iron sulfur-subunit in human liver mitochondria was deduced from cDNA which was isolated by immunoscreening a human liver lambda gtll cDNA library. An isolated clone contains an open reading frame of 786 nucleotides and encodes a mature protein of 252 amino acids with a molecular weight of 28,804. The amino acid sequence was highly homologous with that of bovine heart (94.1%) which has been determined from the purified peptide and that of Escherichia coli sdh B product (50.8%). Striking sequence conservation was found around the three cysteine-rich clusters which have been thought to comprise the iron-sulfur centers of the enzyme. This is the first report on the cDNA sequence of mitochondrial complex II.