scholarly journals Specificity of Alternative Splice Form Detection Using RT-PCR with a Primer Spanning the Exon Junction

BioTechniques ◽  
2003 ◽  
Vol 34 (6) ◽  
pp. 1244-1249 ◽  
Author(s):  
Natalia Shulzhenko ◽  
Anna S. Smirnova ◽  
Andrey Morgun ◽  
Maria Gerbase-DeLima
Keyword(s):  
Alternative Splice ◽  
Splice Form ◽  
Alternative Splice Form ◽  
Rt Pcr ◽  
Exon Junction

An alternative splice form of CMTM8 induces apoptosis

2007 ◽  
Vol 39 (11) ◽  
pp. 2107-2119 ◽  
Author(s):  
Dan Li ◽  
Caining Jin ◽  
Caihua Yin ◽  
Yingmei Zhang ◽  
Bo Pang ◽  
...  
Keyword(s):  
Alternative Splice ◽  
Splice Form ◽  
Alternative Splice Form

scholarly journals An Alternative Splice Form of Mdm2 Induces p53-independent Cell Growth and Tumorigenesis

2003 ◽  
Vol 279 (6) ◽  
pp. 4877-4886 ◽  
Author(s):  
Heather A. Steinman ◽  
Ezra Burstein ◽  
Christopher Lengner ◽  
Joseph Gosselin ◽  
German Pihan ◽  
...  
Keyword(s):  
Cell Growth ◽  
Alternative Splice ◽  
Splice Form ◽  
Alternative Splice Form

scholarly journals Ca2+-ATPase protein expression in mammary tissue

2000 ◽  
Vol 279 (5) ◽  
pp. C1595-C1602 ◽  
Author(s):  
Timothy A. Reinhardt ◽  
Adelaida G. Filoteo ◽  
John T. Penniston ◽  
Ronald L. Horst
Keyword(s):  
Protein Expression ◽  
Milk Fat ◽  
Secretory Pathway ◽  
Apical Membrane ◽  
Mammary Tissue ◽  
Splice Form ◽  
Rt Pcr ◽  
Milk Fat Globule ◽  
Alternatively Spliced ◽  
Fat Globule

Protein expression of plasma membrane Ca2+-ATPases (PMCAs) and the putative Golgi secretory pathway Ca2+-ATPase (SPCA) was examined in rat mammary tissue. As lactation started, PMCA protein expression increased dramatically, and this increased expression paralleled milk production. Mammary PMCA was primarily PMCA2b but was ∼4,000 daltons larger than expected. RT-PCR showed that the primary mammary PMCA2b transcript was alternatively spliced, at splice site A, to include an additional 135 bp, resulting in the insertion of 45 amino acids. This splice form is designated 2bw. PMCA2bw is secreted into milk, associated with the milk fat globule membrane. Therefore, PMCA2bw is located on the apical membrane of the secretory cell. Smaller amounts of PMCA1b and 4b protein were found in mammary tissue. PMCA4b was the major PMCA expressed in developing tissue, and its level declined as lactation started. PMCA1b expression increased moderately during lactation. SPCA protein expression increased 1 wk before parturition and increased further as lactation proceeded. The abundance and cell location of PMCA2b suggest that it is important for macro-Ca2+ homeostasis in lactating tissue. The pattern of expression and abundance of SPCA suggest that it is a candidate for the Golgi Ca2+-ATPase.

scholarly journals Cloning and Functional Analysis of Hypothalamic Homeobox Gene Bsx1a and Its Isoform, Bsx1b

2007 ◽  
Vol 27 (10) ◽  
pp. 3743-3749 ◽  
Author(s):  
Hui-Yi Chu ◽  
Akihira Ohtoshi
Keyword(s):  
Rna Splicing ◽  
Molecular Mechanisms ◽  
Homeobox Gene ◽  
Dna Binding Protein ◽  
Terminal Region ◽  
Splice Form ◽  
Alternative Splice Form ◽  
Transcriptional Reporter ◽  
Reporter Assays ◽  
Immunohistochemical Analyses

ABSTRACT The hypothalamus is a key regulatory unit of the neuroendocrine system and plays an essential role in energy balance and reproduction. Despite its important role, the molecular mechanisms underlying hypothalamic development are not fully understood. Here, we report molecular analyses of a newly identified murine homeobox gene, Bsx/Bsx1a, that is expressed in the developing and postnatal hypothalamus. We demonstrate that BSX1A is a DNA binding protein and a transcriptional activator. Transcriptional reporter assays identified the C-terminal region of BSX1A as an activation domain. We have isolated an alternative splice form of Bsx1a, designated Bsx1b, which retains the N-terminal region but lacks the homeodomain. Analyses of subcellular localization using transfected cell lines revealed that BSX1A and BSX1B localize in the nuclei and cytoplasm, respectively. Immunohistochemical analyses suggested that both BSX1A and BSX1B are expressed in the neonatal hypothalamus. Taking these data together, we propose that alternative RNA splicing is involved in hypothalamic development/function.

scholarly journals BRS1 mediates plant redox regulation and cold responses

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Dongzhi Zhang ◽  
Yuqian Zhao ◽  
Junzhe Wang ◽  
Peng Zhao ◽  
Shengbao Xu
Keyword(s):  
Stress Responses ◽  
Redox Regulation ◽  
Splice Form ◽  
Alternative Splice Form ◽  
Serine Carboxypeptidase ◽  
Over Expression ◽  
Brassinosteroid Signaling ◽  
Cold Responses ◽  
Precise Role

Abstract Background Brassinosteroid-insensitive 1 suppressor 1 (BRS1) is a serine carboxypeptidase that mediates brassinosteroid signaling and participates in multiple developmental processes in Arabidopsis. However, little is known about the precise role of BRS1 in this context. Results In this study, we analyzed transcriptional and proteomic profiles of Arabidopsis seedlings overexpressing BRS1 and found that this gene was involved in both cold stress responses and redox regulation. Further proteomic evidence showed that BRS1 regulated cell redox by indirectly interacting with cytosolic NADP + -dependent isocitrate dehydrogenase (cICDH). One novel alternative splice form of BRS1 was identified in over-expression mutants brs1-1D, which may confer a new role in plant development and stress responses. Conclusions This study highlights the role of BRS1 in plant redox regulation and stress responses, which extends our understanding of extracellular serine carboxypeptidases.

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

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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