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J. Hermans Ch. Rother J. Bichler J. Steppuhn R. G. Herrmann 《Plant molecular biology》1988,10(4):323-330
The nucleotide sequence of the entire nuclear-encoded precursor for subunit delta of the ATP synthase from spinach thylakoid membranes was determined by cDNA sequencing. Appropriate recombinant DNAs were selected from pBR322 and lambda gt11 libraries made from polyadenylated RNA of greening spinach seedlings. The mature protein consists of 187 amino acid residues corresponding to a molecular weight of 20468. The precursor protein (257 amino acid residues; M
r=27676) is probably processed between a Met-Val bond. The predicted secondary structure of the transit sequence (70 residues; 7.2 kDa) resembles that of the Rieske Fe/S polypeptide, but shows little similarity with those of stromal or luminal proteins. The comparison of the chloroplast delta amino acid sequence with the published delta sequences from respiratory ATP synthases of bacterial and mitochondrial sources and from the thylakoid ATP synthase of the cyanobacterium Synechococcus suggests substantial divergence at the genic level although structural elements appear to be remarkably conserved. 相似文献
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J. Steppuhn C. Rother J. Hermans T. Jansen J. Salnikow G. Hauska R. G. Herrmann 《Molecular & general genetics : MGG》1987,210(1):171-177
Summary SummarySeveral cDNA clones encoding the entire Rieske FeS-precursor protein of the chloroplast cytochrome b
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f-complex have been isolated by high density plaque immunoscreening of a phage lambda gt11 cDNA expression library, made from poly A+-RNA of spinach seedlings. The identity of the cDNAs has been confirmed by N-terminal amino acid sequencing of the purified protein. The nucleotide sequence indicates a protein of 247 amino acid residues including a putative transit sequence of 68 amino acids corresponding to molecular masses of 26.3 kDa (precursor) and 18.8 kDa (mature protein; 179 amino acid residues). Alignteins of the sequence with sequences from Rieske FeS-proteins of respiratory electron transport chains, two of bacterial and three of mitochondrial origin, shows little sequence homology, but remarkable similarity in secondary structure including a putative N-terminal transmembrane segment of about 25 residues and the peptides CTHLGCV and CPCHGS in the C-terminal region of the protein that are involved in the binding of the Fe2S2-cluster. 相似文献
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Lars-Gunnar Franzén Gerhard Frank Herbert Zuber Jean-David Rochaix 《Plant molecular biology》1989,12(4):463-474
cDNA clones encoding two Photosystem I subunits of Chlamydomonas reinhardtii with apparent molecular masses of 18 and 11 kDa (thylakoid polypeptides 21 and 30; P21 and P30 respectively) were isolated using oligonucleotides, the sequences of which were deduced from the N-terminal amino acid sequences of the proteins. The cDNAs were sequenced and used to probe Southern and Northern blots. The Southern blot analysis indicates that both proteins are encoded by single-copy genes. The mRNA sizes of the two components are 1400 and 740 nucleotides, respectively. Comparison between the open reading frames of the cDNAs and the N-terminal amino acid sequences of the proteins indicates that the molecular masses of the mature proteins are 17.9 (P21) and 8.1 kDa (P30). Analysis of the deduced protein sequences predicts that both subunits are extrinsic membrane proteins with net positive charges. The amino acid sequences of the transit peptides suggest that P21 and P30 are routed towards the lumenal and stromal sides of the thylakoid membranes, respectively.Abbreviations OEE1, 2 and 3
oxygen evolution enhancer proteins 1, 2 and 3
- Rubisco
ribulose bisphosphate carboxylase/oxygenase
- PS
photosystem
- P21 and P30
C. reinhardtii thylakoid polypeptides 21 and 30 相似文献
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盐藻PSY基因cDNA的克隆、序列测定及其进化分析 总被引:1,自引:0,他引:1
以盐藻总RNA为模板,根据已获得的盐藻PSY基因起始和终止密码子附近的核苷酸序列设计引物,通过RT—PCR扩增获得盐藻PSY cDNA片段.盐藻PSY基因cDNA全长1260bp,编码的盐藻PSY蛋白由420个氨基酸组成,大小为47.3kD,等电点为8.67.在GenBank中进行Blast P检索,发现该片段与许多植物PSY具有较高的相似性(78%~89%).系统进化分析进一步证明,藻类PSY与蓝细菌PSY亲缘关系最近. 相似文献
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我们从重组的人α干扰素处理的单层HeLa细胞常规提取Poly(A)~+RNA作为逆转录合成cDNA第一链之模板,用引物-适配接头法在噬菌质粒pTz19R中构建cDNA文库。以~(32)p-标记的480bpIL-6cDNA片段作探针进行菌落原位及狭缝印迹杂交,筛选出6个阳性克隆。其中两个克隆并用限制酶切分析及DNA序列测定做进一步鉴定。结果证明,一个克隆的cDNA片段长1.3kb,含有人白介素6全长编码区;另一个的cDNA插入片段为0.9kb,缺乏信号肽及成熟IL-6N端30个残基的编码序列。 相似文献
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《Free radical research》2013,47(6):337-346
Superoxide dismutase (SOD: EC 1.15.1.1) catalyzes the dismutation of oxygen radicals and is thought to protect cells against free radical damage. We have isolated and sequenced cDNA clones of the rat Cu/Zn SOD, and have used these clones to map the rat genomic sequences coding for this enzyme. Rat Cu/Zn SOD is coded for by a single copy gene which is transcribed into an mRNA species of approximately 800 bases. Comparison of the nucleotide sequences of rat and human SOD cDNAs shows that they are homologous over 83% of the coding sequences and that in the 3′-untranslated region the extent of homology drops to 66%. The predicted rat SOD amino acid sequence is very similar to that of other eukaryotic SODs, showing 70% homology with the SODs of other mammals. Sequence conservation is particularly high in domains believed to be of functional importance. 相似文献
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菠菜甜菜碱醛脱氢酶基因的克隆和序列分析 总被引:7,自引:0,他引:7
以耐盐的菠菜mRNA为模板,经反转录合成甜菜碱醛脱氢酶(BADH)基因第一链cDNA。在人工合成的两端引物引导下,通过多聚酶链式反应(PCR),扩增获得双链cDNA。把重组有BADH基因的pUC19转化至E.coli DH5α菌株,亚克隆后测定了基因的全序列。所得到的BADH基因全长序列为1491bp,编码497个氨基酸。与文献报道的相比较,核苷酸序列同源性99.8%,氨基酸序列同源性达99.6%。在此基础上,构建了BADH基因的高等植物表达载体。 相似文献
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中国对虾蜕皮抑制激素全长cDNA的克隆及序列分析 总被引:18,自引:1,他引:17
对虾的蜕皮活动由蜕皮抑制激素和蜕皮激素调控,蜕皮抑制激素是甲壳动物CHH家族神经肽的成员之一,通过抑制Y器官蜕皮激素的合成而调节蜕皮,以中国对虾(Fennropenaeus chinensis)眼柄总RNA为材料,采用cDNA末端快速扩增(RACE)方法。首次得到蜕皮抑制激素的全长cDNA(GenBank登录号:AF469187)。该全长cDNA大小为697bp,是由320bp的3′RACE产物和468bp的5′RACE产物拼接而成,Blast搜索结果显示,该全长cDNA与甲壳动物的MIH基因序列具有较高的相似性,用Clustal X进行多序列比较结果表明,由该全长cDNA推导的氨基酸序列与对虾类的MIH的氨基酸序列同源性最高,其中与日本对虾,斑节对虾,刀额新对虾MIH的同源性分别为95.1%,83.1%,79.1%,根据以上数据,推断该697bp的全长cDNA为编码中国对虾MIH前体的cDNA。进一步序列分析表明,编码中国对虾MIH前体cDNA包括312bp的开放阅读框,81bp的3′UTR和302bp的5′UTR;编码103个氨基酸的MIH前体分子包括信号肽和成熟肽,信号肽由28个氨基酸组成,成熟肽由75个氨基酸组成,成熟肽中6个半胱氨酸非常保守。 相似文献
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The primary structure of acetohydroxy acid isomeroreductase from Arabidopsis thaliana was deduced from two overlapping cDNA. The full-length cDNA sequence predicts an amino acid sequence for the protein precursor of 591 residues including a putative transit peptide of 67 amino acids. Comparison of the A. thaliana and spinach acetohydroxy acid isomeroreductases reveals that the sequences are conserved in the mature protein regions, but divergent in the transit peptides and around their putative processing site. 相似文献
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菜花烙铁头蛇毒C-型凝集素基因的克隆与序列分析 总被引:1,自引:1,他引:1
从菜花烙铁头蛇(Trimeresurus jerdonii)的毒腺中提取mRNA,采用RT-PCR技术进行体外扩增,将扩增产物克隆到PMD18-T载体中,最后筛选出一个编码凝集素的基因,命名为TJL。由TJL基因序列推导的氨基酸序列中包含分别由23和135个氨基酸残基组成的信号肽和成熟肽。氨基酸序列比较分析表明,TJL含有半乳糖结合位点和钙离子结合位点,与蝰科蛇毒凝集素TSL、PAL、APL和RSL的同源性较高(87.4%-90.4%),与眼镜蛇科蛇毒凝集素BML的同源性较低(61.5%)。 相似文献
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Two cDNA clones for the barley photosystem I polypeptide which migrates with an apparent molecular mass of 9.5 kDa on SDS-polyacrylamide gels have been isolated using antibodies and an oligonucleotide probe. The determined N-terminal amino acid sequence for the mature polypeptide confirms the identification of the clones. The 644 base-pair sequence of one of the clones contains one large open reading frame coding for a 14 882 Da precursor polypeptide. The molecular mass of the mature polypeptide is 10 193 Da. The hydropathy plot of the polypeptide shows one membrane-spanning region with a predicted -helix secondary structure. The gene for the 9.5 kDa polypeptide has been designated PsaH. 相似文献
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Shyuichiro Inagaki Okitsugu Yamashita 《Archives of insect biochemistry and physiology》1989,10(2):131-139
Complementary (c)DNA coding for an insect yolk protein, the egg-specific protein of the silkworm Bombyx mori was cloned and the nucleotide sequence determined. The sequence covers the entire coding region of 1,677 base pairs with 5′ and 3′ noncoding regions (21 and 115 base pairs, respectively). The deduced amino acid sequence of the egg-specific protein consists of 559 amino acid residues. The NH2-terminal 18 amino acid sequence is enriched in hydrophobic amino acids and assumed to be a signal peptide. A sequence, Asn-X-Thr, a potential N-linked glycosylation site, is found at positions 191 to 193. A serine-rich domain is localized in the region from 63 to 90, in which phosphorylation takes place. Cys His motif in 405 to 415 is analogous to a proposed metal binding sequence. Lys132-Asn133 and Arg228-Asp229 are probably the sites cleaved by the egg-specific protein protease that appears during embryogenesis. The derived amino acid sequence has no appreciable homology to other sequenced proteins. 相似文献
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A cDNA clone encoding a 15.501 Da photosystem I (PSI) subunit of barley was isolated using an oligonucleotide based on the NH2-terminal amino acid sequence of the isolated protein. The polypeptide, which migrates with an apparent molecular mass of 9.5 kDa on denaturing SDS-PAGE, has been designated PSI-N, and the corresponding gene is PsaN. Analysis of the deduced protein sequence indicates a mature protein of 85 amino acid residues and a molecular mass of 9818 Da. PSI-N is a hydrophilic, extrinsic protein with no predicted membrane-spanning regions. The transit peptide of 60 residues (5683 Da) contains a predicted hydrophobic -helix, suggesting that the protein is routed into the thylakoid lumen. Thus, PSI-N is the second known lumenal protein component associated with PSI, together with PSI-F. 相似文献
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在研究狗抗吗啡活性肽PPC过程中,发现它与牛的DBI(diazepambindinginhibitor)的氨基酸序列有很高的同源性,但尚未见到有关狗的DBI的文献报道,为了更好的探讨PPC和DBI相互间的关系,对狗的DBI的cDNA进行了克隆和序列分析。本研究利用大鼠DBI的基因片段为探针,从狗肝脏cDNA文库中,筛选得到了一阳性克隆,并进行了全自动和手工测序,得到了DBI的全长基因。根据EBMLbank序列检索,发现狗的DBI核酸序列与牛的同源性为81%,将其核酸序列翻译成氨基酸序列,进行同源序列比较,结果显示:狗的DBI的氨基酸序列与猪、牛、人、酵母DBI的同源性分别为88.5%、87.4%、83.9%、46.5%。研究还发现狗的DBI序列与抗吗啡活性肽PPCN端62个氨基酸只有两个不同,C端17个氨基酸序列完全相同。只是PPC比DBI中间多了23个氨基酸。 相似文献