Isolation and characterization of a cDNA clone encoding polyubiquitin from the root nodule ofElaeagnus umbellata

1999 ◽  
Vol 77 (9) ◽  
pp. 1270-1278
Author(s):  
Ho Bang Kim ◽  
Chung Sun An
Keyword(s):  
Amino Acids ◽  
In Situ Hybridization ◽  
Amino Acid ◽  
Cdna Clone ◽  
Root Nodule ◽  
Vascular System ◽  
Northern Hybridization ◽  
Elaeagnus Umbellata ◽  
Isolation And Characterization

A cDNA clone encoding polyubiquitin was isolated from a root nodule cDNA library of Elaeagnus umbellata Thunb. by differential hybridization, and its molecular aspects were characterized. The polyubiquitin clone pEuNOD-PUB1 has an insert size of 1642 bp and has the capacity to code for a 458 amino acid residue polyubiquitin protein. The derived amino acid sequence indicates that pEuNOD-PUB1 encodes a polyprotein consisting of six repeats of ubiquitin monomer, except for the last repeat, which has two additional amino acids (Asp-Phe) to be removed in the course of polyubiquitin processing into monomer. The molecular mass of ubiquitin monomer, consisting of 76 amino acids, was predicted to be 8524 Da, and the pI value was predicted to be 7.57. The nucleotide sequence of ubiquitin monomers from pEuNOD-PUB1 showed 73.1-86.0% sequence similarity with those from other organisms. The polyubiquitin mRNA content was four to six times higher in the root nodules than in the leaves and roots. In situ hybridization results showed polyubiquitin transcripts were strongly detected in the meristem zone, in infected cells of the fixation zone, and in the central vascular system. Genomic Southern hybridization revealed that polyubiquitin genes are present as a small multigene family in the genome of E. umbellata.Key words: Elaeagnus umbellata, root nodule, cDNA, polyubiquitin, Northern hybridization, in situ hybridization.

Structures and expression patterns of two cDNA clones encoding S-adenosyl-L-methionine synthetase from the root nodule of Elaeagnus umbellata

2001 ◽  
Vol 28 (9) ◽  
pp. 951
Author(s):  
Sang Ho Lee ◽  
Ho Bang Kim ◽  
Chung Sun An
Keyword(s):  
Amino Acid ◽  
Expression Pattern ◽  
Root Nodule ◽  
Vascular System ◽  
Expression Patterns ◽  
Amino Acid Level ◽  
Nodule Development ◽  
Pcr Analysis ◽  
Cdna Clones ◽  
Elaeagnus Umbellata

This paper originates from an address at the 8th International Symposium on Nitrogen Fixation with Non-Legumes, Sydney, NSW, December 2000 Two cDNA clones encoding S-adenosyl-L-methionine synthetase (SAMS) were isolated from the root nodule cDNA library of Elaeagnus umbellata Thunberg and analysed on the basis of deduced amino acid sequence and expression pattern. Two EuSAMS clones shared 75–84% identity at the nucleotide level, and 85–95% identity at the amino acid level, with the other plant SAMS genes. Genomic Southern hybridization revealed the presence of more than two copies of SAMS genes in the genome of E. umbellata. Reverse transcriptase-mediated polymerase chain reaction (RT–PCR) analysis showed EuSAMS1 transcripts were more abundant than those of EuSAMS2. Similar to the expression pattern of other plant SAMS genes, both genes were expressed at higher levels in root than in leaf. During nodule development, expression of both genes was increased, with the highest level at 6–8 week after inoculation, and decreased rapidly thereafter. In situ hybridization analysis also showed both SAMS transcripts in the meristem zone, the infected cells of the fixation zone and in the central vascular system of root nodules. However, EuSAMS2 transcripts were strongly detected in the prefixation zone, whereas EuSAMS1 transcripts were hardly detected. These results suggest different regulatory mechanisms for the two genes in the root nodule. The expression pattern of SAMS genes in the root nodule may correlate mostly with cell wall synthesis, polyamine biosynthesis and other methylation-mediated functions.

scholarly journals Isolation and Characterization of a Novel Member of the Relaxin/Insulin Family from the Testis of the Frog Rana esculenta*

Endocrinology ◽  
2001 ◽  
Vol 142 (7) ◽  
pp. 3231-3238 ◽  
Author(s):  
Gianluca de Rienzo ◽  
Francesco Aniello ◽  
Margherita Branno ◽  
Sergio Minucci
Keyword(s):  
In Situ Hybridization ◽  
Amino Acid ◽  
Messenger Rna ◽  
Rana Esculenta ◽  
Amino Acid Sequences ◽  
Characteristic Functional ◽  
Isolation And Characterization ◽  
Single Transcript ◽  
A Domains

Abstract A complementary DNA (cDNA) encoding a frog relaxin/insulin member family (fRLX) from testis cDNA library was isolated and characterized. The fRLX cDNA predicted a 155-amino acid protein with a low homology to mammalian RLF and relaxin. Northern blot analysis revealed a single transcript expressed in the interstitial compartment, RT-PCR, evidenced that fRLX is expressed at low levels in the oviduct and ovary too. The predicted mature fRLX protein, composed of the signal peptide, B, C, and A domains, has conserved amino acid sequences in the characteristic functional domains. A different expression of the transcript was found during the frog reproductive cycle, with a peak in Spring. After administration of ethane dimethane sulfonate, by in situ hybridization, fRLX messenger RNA disappeared from the interstitial compartment and reappeared again at the time of generating of a new population of Leydig cells (LC), strongly indicating that LC are the interstitial cell type expressing fRLX. Preliminary results obtained by in situ hybridization, performed on testis of hypophysectomized frogs evidenced a pituitary control of fRLX expression. This study is the first cloning of a relaxin/insulin family member in a nonmammalian vertebrate. In addition, because fRLX expression changes during the annual cycle suggesting its involvement in spermatogenesis, fRLX may be considered a new marker for the study of spermatogenesis in the Rana esculenta.

A facile one pot route for the synthesis of imide tethered peptidomimetics

2016 ◽  
Vol 14 (2) ◽  
pp. 556-563 ◽  
Author(s):  
Veladi Panduranga ◽  
Girish Prabhu ◽  
Roopesh Kumar ◽  
Basavaprabhu Basavaprabhu ◽  
Vommina V. Sureshbabu
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Efficient Method ◽  
One Pot ◽  
Protected Amino Acid

A simple and efficient method for the synthesis of N,N’-orthogonally protected imide tethered peptidomimetics is presented. The imide peptidomimetics were synthesized by coupling the in situ generated selenocarboxylate of Nα-protected amino acids with Nα-protected amino acid azides in good yields.

Isolation and characterization of some novel genes of the apolipoprotein A-I family in Japanese eel, Anguilla japonica

2011 ◽  
Vol 6 (4) ◽  
pp. 545-557 ◽  
Author(s):  
Malay Choudhury ◽  
Takahiro Oku ◽  
Shoji Yamada ◽  
Masaharu Komatsu ◽  
Keita Kudoh ◽  
...  
Keyword(s):  
Amino Acids ◽  
Molecular Weight ◽  
Amino Acid ◽  
Consensus Sequence ◽  
Structural Features ◽  
Lipid Binding ◽  
Japanese Eel ◽  
Amino Acid Sequences ◽  
Novel Genes ◽  
Isolation And Characterization

AbstractApolipoproteins such as apolipoprotein (apo) A-I, apoA-IV, and apoE are lipid binding proteins synthesized mainly in the liver and the intestine and play an important role in the transfer of exogenous or endogenous lipids through the circulatory system. To investigate the mechanism of lipid transport in fish, we have isolated some novel genes of the apoA-I family, apoIA-I (apoA-I isoform) 1–11, from Japanese eel by PCR amplification. Some of the isolated genes of apoIA-I corresponded to 28kDa-1 cDNAs which had already been deposited into the database and encoded an apolipoprotein with molecular weight of 28 kDa in the LDL, whereas others seemed to be novel genes. The structural organization of all apoIA-Is consisted of four exons separated by three introns. ApoIA-I10 had a total length of 3232 bp, whereas other genes except for apoIA-I9 ranged from 1280 to 1441 bp. The sequences of apoIA-Is at the exon-intron junctions were mostly consistent with the consensus sequence (GT/AG) at exon-intron boundaries, whereas the sequences of 3′ splice acceptor in intron 1 of apoIA-I1-7 were (AC) but not (AG). The deduced amino acid sequences of all apoIA-Is contained a putative signal peptide and a propeptide of 17 and 5 amino acid residues, respectively. The mature proteins of apoIA-I1-3, 7, and 8 consisted of 237 amino acids, whereas those of apoIA-I4-6 consisted of 239 amino acids. The mature apoIA-I10 sequence showed 65% identity to amino acid sequence of apoIA-I11 which was associated with an apolipoprotein with molecular weight of 23 kDa in the VLDL. All these mature apoIA-I sequences satisfied the common structural features depicted for the exchangeable apolipoproteins such as apoA-I, apoA-IV, and apoE but apoIA-I11 lacked internal repeats 7, 8, and 9 when compared with other members of apoA-I family. Phylogenetic analysis showed that these novel apoIA-Is isolated from Japanese eel were much closer to apoA-I than apoA-IV and apoE, suggesting new members of the apoA-I family.

scholarly journals Lotus japonicus Contains Two Distinct ENOD40 Genes That Are Expressed in Symbiotic, Nonsymbiotic, and Embryonic Tissues

2000 ◽  
Vol 13 (9) ◽  
pp. 987-994 ◽  
Author(s):  
Emmanouil Flemetakis ◽  
Nektarios Kavroulakis ◽  
Nicolette E. M. Quaedvlieg ◽  
Herman P. Spaink ◽  
Maria Dimou ◽  
...  
Keyword(s):  
Root Nodule ◽  
Vascular System ◽  
Lotus Japonicus ◽  
Mrna Accumulation ◽  
Mesorhizobium Loti ◽  
Embryonic Tissues ◽  
Early Nodulin ◽  
Gene Transcripts ◽  
Young Seed

ENOD40, an early nodulin gene, has been postulated to play a significant role in legume root nodule ontogenesis. We have isolated two distinct ENOD40 genes from Lotus japonicus. The transcribed regions of the two ENOD40 genes share 65% homology, while the two promoters showed no significant homology. Both transcripts encode a putative dodecapeptide similar to that identified in other legumes forming determinate nodules. Both ENOD40 genes are coordinately expressed following inoculation of roots with Mesorhizobium loti or treatment with purified Nod factors. In the former case, mRNA accumulation could be detected up to 10 days following inoculation while in the latter case the accumulation was transient. High levels of both ENOD40 gene transcripts were found in nonsymbiotic tissues such as stems, fully developed flowers, green seed pods, and hypocotyls. A relatively lower level of both transcripts was observed in leaves, roots, and cotyledons. In situ hybridization studies revealed that, in mature nodules, transcripts of both ENOD40 genes accumulate in the nodule vascular system; additionally, in young seed pods strong signal is observed in the ovule, particularly in the phloem and epithelium, as well as in globular stage embryos.

Uptake and metabolism of amino acids by the dog liver perfused in situ

1962 ◽  
Vol 202 (3) ◽  
pp. 407-414 ◽  
Author(s):  
Rapier H. McMenamy ◽  
William C. Shoemaker ◽  
Jonas E. Richmond ◽  
David Elwyn
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Mixture ◽  
Acid Mixture ◽  
Sharp Rise ◽  
Dog Blood ◽  
Dog Liver ◽  
Amino Acid Concentrations ◽  
Uptake And Metabolism

Dog livers were perfused in situ for periods up to 6 hr with dog blood recycled through a pump-oxygenator. An amino acid mixture was administered for 90 min. Concentrations of amino acids were determined at intervals of 30 min or more. Rates of uptake and metabolism were calculated. After the start of perfusion, there is a fall in most plasma amino acid concentrations and a reciprocal rise in liver amino acids. Addition of amino acids causes a sharp rise in plasma amino acids. There is a rapid uptake of most of the amino acids by liver, although the concentrations of amino acids in liver fail to rise appreciably. Notable exceptions are valine, leucine, and isoleucine. Uptake of amino acids stimulates: a) an increase in the rate of synthesis of urea which ultimately accounts for 90% of the metabolized amino acids; b) a net synthesis of ornithine; and c) net noncatabolic metabolism of amino acids which may in part be protein synthesis. The results support the view that the liver temporarily stores a part of ingested amino acids as proteins, and subsequently makes them available to other organs.

Amino acid transport by juxtamedullary nephrons: distal reabsorption and recycling

1988 ◽  
Vol 255 (3) ◽  
pp. F397-F407 ◽  
Author(s):  
W. H. Dantzler ◽  
S. Silbernagl
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Transport ◽  
Acid Transport ◽  
Acidic Amino Acids ◽  
Vasa Recta ◽  
Collecting Ducts ◽  
Distal Site

Amino acid transport by juxtamedullary (JM) nephrons and its relationship to transport by superficial cortical (SC) nephrons and to function of vasa recta and collecting ducts were examined in vivo and in situ by free-flow micropuncture of Henle's loops, collecting ducts, and vasa recta and by continuous microinfusion of Henle's loops in exposed rat papillae. Fractional deliveries (FDs) of six neutral amino acids, two acidic amino acids, and taurine to tips of Henle's loops of JM nephrons could be substantially below those to early distal loops of SC nephrons, indicating that reabsorption before loop tips could be greater in JM than in SC nephrons. FDs to collecting ducts lower than to JM loop tips suggested reabsorption distal to loop tips. This was confirmed by continuous microinfusion of ascending limbs of Henle's loops. Distal site of reabsorption is unknown, but amino acids may move passively out of the thin ascending limb and be recycled into vasa recta and descending limb. Recycling of amino acids was supported by high FDs to tips of Henle's loops (sometimes greater than 1.0), higher concentrations in ascending than in descending vasa recta at same papilla level, and high mean concentrations in vasa recta.

Isolation and characterization of a zebra finch aromatase cDNA: in situ hybridization reveals high aromatase expression in brain

1994 ◽  
Vol 24 (1-4) ◽  
pp. 227-237 ◽  
Author(s):  
P. Shen ◽  
C.W. Campagnoni ◽  
K. Kampf ◽  
B.A. Schlinger ◽  
A.P. Arnold ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Zebra Finch ◽  
Aromatase Expression ◽  
Isolation And Characterization
Sign in / Sign up

Export Citation Format

Share Document