Temporal classification and mapping of non-polyadenylated transcripts of an invertebrate iridovirus

The temporal expression of the 54 Chilo iridescent virus (CIV) virion protein genes was investigated by combining drug treatments that inhibit protein or DNA synthesis and an RT-PCR strategy particularly suitable for non-polyadenylated mRNAs. This method generates a uniform 3′ terminus by ligation of a 5′-phosphorylated oligonucleotide to the 3′ end of the transcript that is recognized by a complementary primer during RT-PCR. This analysis showed that CIV virion proteins are encoded by genes in all three predetermined temporal classes: 23 immediate-early, 11 delayed-early and seven late virion gene transcripts were identified and assigned to ORFs. Early transcription of many virion protein genes supports the notion that virion proteins may also play essential roles in the initial stages of infection. In addition, some of the early gene products present in the virion may reflect the intracellular path that the virus follows during infection.

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Detection of Nucleic Acids in Cells or Tissue Sections Using In situ Polymerase Chain Reaction (PCR)

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100162545 ◽

1996 ◽

Vol 54 ◽

pp. 16-17

Author(s):

J. R. Hully ◽

K. R. Luehrsen ◽

K. Aoyagi ◽

C. Shoemaker ◽

R. Abramson

Keyword(s):

Nucleic Acids ◽

Viral Infections ◽

Anatomical Location ◽

Rt Pcr ◽

Reverse Transcription Pcr ◽

Cellular Source ◽

Gene Transcripts ◽

Initial Description ◽

In Cells

The development of PCR technology has greatly accelerated medical research at the genetic and molecular levels. Until recently, the inherent sensitivity of this technique has been limited to isolated preparations of nucleic acids which lack or at best have limited morphological information. With the obvious exception of cell lines, traditional PCR or reverse transcription-PCR (RT-PCR) cannot identify the cellular source of the amplified product. In contrast, in situ hybridization (ISH) by definition, defines the anatomical location of a gene and/or it’s product. However, this technique lacks the sensitivity of PCR and cannot routinely detect less than 10 to 20 copies per cell. Consequently, the localization of rare transcripts, latent viral infections, foreign or altered genes cannot be identified by this technique. In situ PCR or in situ RT-PCR is a combination of the two techniques, exploiting the sensitivity of PCR and the anatomical definition provided by ISH. Since it’s initial description considerable advances have been made in the application of in situ PCR, improvements in protocols, and the development of hardware dedicated to in situ PCR using conventional microscope slides. Our understanding of the importance of viral latency or viral burden in regards to HIV, HPV, and KSHV infections has benefited from this technique, enabling detection of single viral copies in cells or tissue otherwise thought to be normal. Clearly, this technique will be useful tool in pathobiology especially carcinogenesis, gene therapy and manipulations, the study of rare gene transcripts, and forensics.

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Faculty Opinions recommendation of Molecular interpretation of ERK signal duration by immediate early gene products.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1002477.109609 ◽

2002 ◽

Author(s):

Carlos Ibáñez

Keyword(s):

Immediate Early Gene ◽

Early Gene ◽

Immediate Early ◽

Signal Duration ◽

Gene Products ◽

Molecular Interpretation

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Differential effects of human recombinant interferons on the expression of two early gene products of Epstein-Barr virus

Antiviral Research ◽

10.1016/0166-3542(92)90092-j ◽

1992 ◽

Vol 17 (1) ◽

pp. 79-89 ◽

Cited By ~ 4

Author(s):

Bodil Lidin ◽

Eddie W. Lamon

Keyword(s):

Epstein Barr Virus ◽

Early Gene ◽

Gene Products ◽

Differential Effects ◽

Barr Virus ◽

Epstein Barr

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Influence of estradiol on bovine trophectoderm and uterine gene transcripts around maternal recognition of pregnancy

Biology of Reproduction ◽

10.1093/biolre/ioab091 ◽

2021 ◽

Author(s):

Emmalee J Northrop-Albrecht ◽

Jerica J J Rich ◽

Robert A Cushman ◽

Runan Yao ◽

Xijin Ge ◽

...

Keyword(s):

Artificial Insemination ◽

Mrna Level ◽

Fixed Time ◽

Beef Cows ◽

Differentially Expressed ◽

Differentially Expressed Proteins ◽

Rt Pcr ◽

Embryo Survival ◽

Uterine Fluid ◽

Gene Transcripts

Abstract Embryo survival and pregnancy success is increased among animals that exhibit estrus prior to fixed time artificial insemination (AI), but there are no differences in conceptus survival to d16. The objective of this study was to determine effects of preovulatory estradiol on uterine transcriptomes, select trophectoderm transcripts, and uterine luminal fluid (ULF) proteins. Beef cows/heifers were synchronized, artificially inseminated (d0), and grouped into either high (highE2) or low (lowE2) preovulatory estradiol. Uteri were flushed (d16); conceptuses and endometrial biopsies (n = 29) were collected. RNA sequencing was performed on endometrium. Real-Time PCR (RT-PCR) was performed on trophectoderm (TE; n = 21) RNA to measure relative abundance of IFNT, PTGS2, TM4SF1, C3, FGFR2, and GAPDH. Uterine fluid was analyzed using 2D LC–MS/MS based iTRAQ method. RT-PCR data were analyzed using the MIXED procedure in SAS. There were no differences in mRNA abundances in TE, but there were 432 differentially expressed genes (DEGs) (253 downregulated, 179 upregulated) in highE2/conceptus versus lowE2/conceptus groups. There were also 48 differentially expressed proteins (DEPs; 19 upregulated, 29 downregulated), 6 of these were differentially expressed (FDR < 0.10) at the mRNA level. Similar pathways for mRNA and proteins included: calcium signaling, protein kinase A signaling, and corticotropin releasing hormone (CRH) signaling. These differences in uterine function, may be preparing the conceptus for improved likelihood of survival after d16 among highE2 animals.

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Chilo iridescent virus encodes two functional metalloproteases

Archives of Virology ◽

10.1007/s00705-018-4108-z ◽

2018 ◽

Vol 164 (3) ◽

pp. 657-665 ◽

Cited By ~ 1

Author(s):

Aydın Yesilyurt ◽

Hacer Muratoglu ◽

Zihni Demirbag ◽

Remziye Nalcacioglu

Keyword(s):

Chilo Iridescent Virus ◽

Iridescent Virus

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Capacitative Ca2+ entry in agonist-induced pulmonary vasoconstriction

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.2001.280.5.l870 ◽

2001 ◽

Vol 280 (5) ◽

pp. L870-L880 ◽

Cited By ~ 111

Author(s):

Sharon S. McDaniel ◽

Oleksandr Platoshyn ◽

Jian Wang ◽

Ying Yu ◽

Michele Sweeney ◽

...

Keyword(s):

Transient Receptor Potential ◽

Channel Blocker ◽

Receptor Potential ◽

Rt Pcr ◽

Pulmonary Vasoconstriction ◽

Store Depletion ◽

Intracellular Ca ◽

Gene Transcripts ◽

Transient Receptor ◽

Sustained Increase

Agonist-induced increases in cytosolic Ca2+ concentration ([Ca2+]cyt) in pulmonary artery (PA) smooth muscle cells (SMCs) consist of a transient Ca2+ release from intracellular stores followed by a sustained Ca2+ influx. Depletion of intracellular Ca2+ stores triggers capacitative Ca2+ entry (CCE), which contributes to the sustained increase in [Ca2+]cyt and the refilling of Ca2+ into the stores. In isolated PAs superfused with Ca2+-free solution, phenylephrine induced a transient contraction, apparently by a rise in [Ca2+]cyt due to Ca2+ release from the intracellular stores. The transient contraction lasted for 3–4 min until the Ca2+ store was depleted. Restoration of extracellular Ca2+ in the presence of phentolamine produced a contraction potentially due to a rise in [Ca2+]cyt via CCE. The store-operated Ca2+ channel blocker Ni2+ reduced the store depletion-activated Ca2+ currents, decreased CCE, and inhibited the CCE-mediated contraction. In single PASMCs, we identified, using RT-PCR, five transient receptor potential gene transcripts. These results suggest that CCE, potentially through transient receptor potential-encoded Ca2+ channels, plays an important role in agonist-mediated PA contraction.

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