Ligation-mediated Molecular Analysis of Influenza Subtypes, Splicing and Protein Glycosylation

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Conze, Tim,1977-Uppsala universitet,Institutionen för genetik och patologi,Molekylära verktyg,Landegren (författare)

Ligation-mediated Molecular Analysis of Influenza Subtypes, Splicing and Protein Glycosylation

BokEngelska2010

Förlag, utgivningsår, omfång ...

Uppsala :Acta Universitatis Upsaliensis,2010
39 s.
electronicrdacarrier

Nummerbeteckningar

LIBRIS-ID:oai:DiVA.org:uu-120016
ISBN:9789155477431
https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-120016URI

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Språk:engelska
Sammanfattning på:engelska

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Ämneskategori:vet swepub-contenttype
Ämneskategori:dok swepub-publicationtype

Serie

Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine,1651-6206 ;532

Anmärkningar

Binder-based assays are employed throughout the life sciences. Powerful signal amplification techniques have enabled detection of very rare molecule species diluted in simple buffers. Unspecific binding of primary binders leads to increased background in more complex samples. By requiring two recognition events, ligation-based molecular analyses provide highly specific detection of biomolecules in complex samples. We developed a highly multiplexed padlock-ligation assay targeting signature sequences in the hemagglutinin and neuraminidase genes. From a panel of 77 avian influenza isolates of all major serotypes, 97% were genotyped correctly in accordance with previous classifications by classical diagnostic methods (Paper I). Alternative splicing is an important mechanism expanding the proteome. Current analysis techniques fail to provide sequences of complete transcripts beyond the read length of sequencing instruments. We devised and implemented a strategy to compress the sequence information contained in the splicing pattern of a transcript into the presence or absence of sequence-blocks. We demonstrate that this assay yields information about the splicing patterns in thousands of transcripts from cellular cDNA (Paper II). Expression changes of mucin proteins and glycosylation structures are frequently observed from the early stages of cancer development. Expression of mucin 2 and sialyl-Tn are common features of intestinal metaplasia and gastric cancer, and are known to co-locate. Here we have developed an in situ proximity ligation assay (PLA) directed against mucin 2 and sialyl-Tn. Our study on intestinal metaplasia and gastric cancer tissue sections identified mucin 2 as a major carrier of sialyl-Tn in these conditions, and demonstrated how conveniently glycosylation of proteins can be studied by in situ PLA (Paper III). This thesis shows how the dual recognition requirement of ligation-based assays can be employed to detect target molecules with high specificity, to analyze several sequence features of nucleic acids or to study the proximity of two antigens in situ.

Ämnesord och genrebeteckningar

MEDICIN OCH HÄLSOVETENSKAP Medicinska och farmaceutiska grundvetenskaper Medicinsk genetik hsv//swe
MEDICAL AND HEALTH SCIENCES Basic Medicine Medical Genetics hsv//eng
ligase
proximity ligation
gastric cancer
glycosylation
alternative splicing
avian influenza
padlock probe
Molecular medicine (genetics and pathology)
Molekylär medicin (genetik och patologi)
Molekylär medicin
Molecular Medicine

Biuppslag (personer, institutioner, konferenser, titlar ...)

Landegren, Ulf,ProfUppsala universitet,Institutionen för genetik och patologi (preses)
Nilsson, Mats,ProfUppsala universitet,Institutionen för genetik och patologi (preses)
Söderberg, Ola,DocentUppsala universitet,Institutionen för genetik och patologi (preses)
Glökler, Jörn,Dr.Max Planck Institute for Molecular Genetics (opponent)
Uppsala universitetInstitutionen för genetik och patologi (creator_code:org_t)

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Av författaren/redakt...: Conze, Tim, 1977 ...; Landegren, Ulf, ...; Nilsson, Mats, P ...; Söderberg, Ola, ...; Glökler, Jörn, D ...

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