Molecular Cloning and Characterization of the Pig Homologue to Human Cd29, the Integrin ??1 Subunit1

Jiménez-Marín, Ángeles; Jj, Garrido; Df, de Andrés-Cara; Morera, L.; Mj, Barbancho; Llanes, D.

doi:10.1097/00007890-200008270-00019

Cited by 16 publications

(14 citation statements)

References 26 publications

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“…Punctual changes between human and swine CD29 molecules within the ligand binding domain, and/or the regulatory domain suggest potential differences between human and porcine CD29 relative to the human CD29 ligand [94]. CD29 mRNA were expressed in a variety of porcine tissues with different intensities [95] and CD29 integrin is widely distributed and found on all the cell lines tested in one study [91].…”

Section: The 1 Integrin Subfamily Cd49 A-f/cd29 Heterodimers: the Vlmentioning

confidence: 95%

Membrane markers of the immune cells in swine: an update

Piriou-Guzylack¹,

2008

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-Besides their breeding value, swine are increasingly used as biomedical models. As reported in three international swine clusters of differentiation (CD) workshops and in the animal homologue section of the last workshop for the determination of human leukocyte differentiation antigens (HLDA 8), characterisation of leukocyte surface antigens by monoclonal antibodies and other molecular studies have determined the cell lineages and blood leukocyte subsets implicated in the immune response, including cell adhesion molecules involved in cell trafficking. This review focusses on the current state of knowledge of porcine leukocyte differentiation and major histocompatibility complex (SLA) molecules. Examples of porcine particularities such as the double-positive T lymphocytes with the phenotype CD4 + CD8 low and CD4 − CD8 low T cell subsets and the persistence of SLA class II after T-lymphocyte activation are illustrated, as well as the shared characteristics of the Artiodactyla group, such as the high proportion of TcR (T cell receptor) T cells in blood and other lymphoid tissues. Furthermore, discrepancies between swine and humans, such as CD16 expression on dendritic cells and CD11b (wCD11R1) tissue distribution are outlined. The rapidly growing information should facilitate manipulation of the swine immune system towards improving disease control, and open new avenues for biomedical research using the pig as a model. cluster of differentiation (CD) / monoclonal antibody / immune system / histocompatibility system / pig

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Section: The 1 Integrin Subfamily Cd49 A-f/cd29 Heterodimers: the Vlmentioning

confidence: 95%

Membrane markers of the immune cells in swine: an update

Piriou-Guzylack¹,

2008

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“…A monoclonal antibody, GE2B6, against rpCD41-F2R2 and two polyclonal antibodies, antirpCD41-F1R1 and anti-rpCD41-F2R2, were produced using previously described immunization and cells fusion procedures (Arce et al, 2002;Jiménez-Marín et al, 2000). Briefly, female BALB/c mice were immunized with 50 µg of rpCD51.…”

Section: Antibodies Productionmentioning

confidence: 99%

“…PCR product was ligated into the expression vector pET28b and used to transform Escherichia coli strain BL21 (DE3) (Novagen). Recombinant proteins (rpCD41-F1R1 and rpCD41-F2R2), expression and purification were carried out following previously procedures described by us (Jiménez-Marín et al, 2000). …”

mentioning

confidence: 99%

Molecular Cloning, Characterization, Expression Analysis and Chromosomal Localization of the Gene Coding for the Porcine αIIb Subunit of the αIIbβ3 Integrin Platelet Receptor

Esteso¹,

Jiménez-Marín²,

Sanz³

et al. 2011

Molecular Cloning - Selected Applications in Medicine and Biology

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Molecular Cloning, Characterization, Expression Analysis and Chromosomal Localization of the Gene Coding for the Porcine αIIb Subunit of the αIIbβ3 Integrin Platelet Receptor 111 RNA isolation, RT-PCR and RACETotal RNA from platelets, cells or tissues was purified according to the M-MLV Reverse Transcriptase system (Invitrogene) using the random primers pd(N) 6 -5'-PO 3 NA + Salt (Pharmacia Biotech). RNA samples were kept at -80ºC after controlling the quality on a denaturing agarose gel. 5 µg RNA, resuspended in 9.5 µl water, were heated for 3 min at 65ºC in the presence of random hexamers (7.5 µM final concentration), and then cooled in ice. RNA was reverse transcribed using 1 µl Moloney murine leukemia virus reverse transcriptase (200 units/µl) (GibcoBRL) for 1 h at 42ºC in a final volume of 20 µl containing 4 µl of 5X reverse transcriptase buffer, 0,5 µl ribonuclease inhibitor (50 U/µl) (Roche), 1 µl 20 mM dNTP (Pharmacia) and 2 µl 0.1 M dithiothreitol. After 10 min at room temperature, 1 h at 42 º C, and 10 min at 95ºC, DEPC H 2 O were added until a final volume of 100 µl. 2 µl of this mixture were subjected to PCR using 1µl Tth DNA polymerase (1U/µl) (Biotools) and 2,5 µl each CD41-specific primer (20µM) (see Table 1) in a final volume of 50 µl containing 5 µl 10x buffer, 2 µl MgCl 2 (50 mM), and 8 µl dNTP MIX (1,25 mM each) (Biotools). The amplification consisted in 35 cycles of PCR and each cycle consisted of incubations at 94ºC for 1 min, TmºC for 1 min, and 72ºC for 1 min. The amplifications were electrophoresed on 1% agarose/1X TAE gel. RT-PCR on RNA18S cDNA was used as a control. For RACE (Rapid Amplification of cDNA Ends), 1 µg total RNA from platelet was used to reverse-transcribe using 1 µl Moloney murine leukemia virus reverse transcriptase (200 units/µl) (GibcoBRL) for 1 h at 42ºC in a final volume of 20 µl containing 4 µl of 10X reverse transcriptase buffer, 1,0 µl ribonuclease inhibitor (50 U/µl) (Roche), 4 µl 2,5 mM dNTP (Pharmacia) and 2 µl 3' RACE ADAPTER (20 µM) in a final volume of 20 µL. 3' CD41 cDNAs were obtained by PCR using a specific porcine CD41 primer and the anchor primer provided in the kit (Table 1). DNA sequencing and sequences analysisSequencing was performed using ABI PRISM BigDye Terminator Cycle Sequencing Ready Reaction Kit (Applied Biosystems, Foster City, CA, USA) on a thermal DNA cycler GeneAmp PCR System 2400 (Applied Biosystems, Foster City, CA, USA), according to the instructions of the manufacturer, and analysed on an ABI PRISM 3100 Sequencer (Applied Biosystems, Foster City, CA, USA). Porcine CD41 sequence has been deposited at GenBank under number JF808665. Sequences were analyzed using the analysis software LaserGene (DNAstar, Londo, UK) and the analysis tools provided by the expasy web site . Primers design was performed with Oligo 6 (MBI, Cascade, CO, USA) and Amplify 3 (). Multiple alignment among CD41 peptide sequences from Sus scrofa (GenBank accession no. JF808665 ), Homo sapiens (GenBank...

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“…The selected genes were validated by RT-qPCR analysis to confirm the result of gene expression level with microarray data and the result of gene expression level of four expressed genes was agreed with the DNA microarray expression data (Table 1). There were Na + /K + transporting ATPase (ATP1B1) [14], H3 histone (H3F3A) [15], integrin beta-1 subunit (ITGB1) [16] and rhodopsin (RHO) [17]. They were up-regulated genes in ANP-stimulated LLC-PK1 cells and have been found to have the correlation with the renal protective system.…”

Section: Rt-qpcr Validation Of Array Analysismentioning

confidence: 99%

Gene expression analysis in LLC-PK1 renal tubular cells by atrial natriuretic peptide (ANP): correlation of homologous human genes with renal response

et al. 2007

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We used human DNA microarray to explore the differential gene expression profiling of atrial natriuretic peptide (ANP)-stimulated renal tubular epithelial kidney cells (LLC-PK1) in order to understand the biological effect of ANP on renal kidney cell's response. Gene expression profiling revealed 807 differentially expressed genes, consisting of 483 up-regulated and 324 down-regulated genes. The bioinformatics tool was used to gain a better understanding of differentially expressed genes in porcine genome homologous with human genome and to search the gene ontology and category classification, such as cellular component, molecular function and biological process. Four up-regulated genes of ATP1B1, H3F3A, ITGB1 and RHO that were typically validated by real-time quantitative PCR (RT-qPCR) analysis serve important roles in the alleviation of renal hypertrophy as well as other related effects. Therefore, the human array can be used for gene expression analysis in pig kidney cells and we believe that our findings of differentially expressed genes served as genetic markers and biological functions can lead to a better understanding of ANP action on the renal protective system and may be used for further therapeutic application.

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Molecular Cloning and Characterization of the Pig Homologue to Human Cd29, the Integrin ??1 Subunit1

Cited by 16 publications

References 26 publications

Membrane markers of the immune cells in swine: an update

Membrane markers of the immune cells in swine: an update

Molecular Cloning, Characterization, Expression Analysis and Chromosomal Localization of the Gene Coding for the Porcine αIIb Subunit of the αIIbβ3 Integrin Platelet Receptor

Gene expression analysis in LLC-PK1 renal tubular cells by atrial natriuretic peptide (ANP): correlation of homologous human genes with renal response

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