In situPCR on Plant Material with Sub-cellular Resolution

Bo Johansen

doi:10.1006/anbo.1997.0502

In situPCR on Plant Material with Sub-cellular Resolution

Bo Johansen

Københavns Universitet

Publikation: Bidrag til tidsskrift › Tidsskriftsartikel › Forskning › peer review

Abstract

In situPCR and reverse transcribedin situPCR have been tested on leaves of sugar cane fixed in FAA, 4% PFA or 2% PFA+2.5% GA.In situPCR amplification of the gene coding for ribulose-1,5-bisphosphate carboxylase/oxygenase (rbcL) was successfully performed following all three fixation protocols. As expected the PCR product was restricted to the plastids of all cells. Reverse transcribedin situPCR was performed onrbcL mRNA and in this case the PCR product was restricted to the plastids of the bundle sheath cells. This is the first report ofin situPCR on plant material and only the second report ofin situPCR with sub-cellular resolution.In situPCR onrbcL may prove to be a valuable positive control for futurein situPCR studies on plant material.Copyright 1997 Annals of Botany Company 10.1006/anbo.1997.0502

Originalsprog	Engelsk
Tidsskrift	Annals of Botany
Vol/bind	80
Udgave nummer	5
Sider (fra-til)	697-700
Antal sider	4
ISSN	0305-7364
DOI	https://doi.org/10.1006/anbo.1997.0502
Status	Udgivet - nov. 1997
Udgivet eksternt	Ja

Emneord

c -plants
in situ pcr
pcr
rbc l
reversed transcribed in situ

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title = "In situPCR on Plant Material with Sub-cellular Resolution",

abstract = "In situ PCR and reverse transcribed in situ PCR have been tested on leaves of sugar cane fixed in FAA, 4\% PFA or 2\% PFA+2.5\% GA. In situ PCR amplification of the gene coding for ribulose-1,5-bisphosphate carboxylase/oxygenase (rbcL) was successfully performed following all three fixation protocols. As expected the PCR product was restricted to the plastids of all cells. Reverse transcribed in situ PCR was performed on rbcL mRNA and in this case the PCR product was restricted to the plastids of the bundle sheath cells. This is the first report of in situ PCR on plant material and only the second report of in situ PCR with sub-cellular resolution. In situ PCR on rbcL may prove to be a valuable positive control for future in situ PCR studies on plant material.",

keywords = "c -plants, in situ pcr, pcr, rbc l, reversed transcribed in situ",

author = "Bo Johansen",

note = "Funding Information: This research was supported by grants from the Danish Natural Science Research Council (9503576 and 11-1030-1). Dr V. Albert kindly made comments on the manuscript. I thank Mrs Charlotte Hansen and Mrs Kate Jensen for technical assistance and Mr Flemming Sarup for photographic work. The stimulating support from {\textquoteleft}Nedl{\o}bsr{\o}ret{\textquoteright} is gratefully acknowledged.",

year = "1997",

month = nov,

doi = "10.1006/anbo.1997.0502",

language = "English",

volume = "80",

pages = "697--700",

journal = "Annals of Botany",

issn = "0305-7364",

publisher = "Oxford University Press",

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T1 - In situPCR on Plant Material with Sub-cellular Resolution

AU - Johansen, Bo

N1 - Funding Information: This research was supported by grants from the Danish Natural Science Research Council (9503576 and 11-1030-1). Dr V. Albert kindly made comments on the manuscript. I thank Mrs Charlotte Hansen and Mrs Kate Jensen for technical assistance and Mr Flemming Sarup for photographic work. The stimulating support from ‘Nedløbsrøret’ is gratefully acknowledged.

PY - 1997/11

Y1 - 1997/11

N2 - In situ PCR and reverse transcribed in situ PCR have been tested on leaves of sugar cane fixed in FAA, 4% PFA or 2% PFA+2.5% GA. In situ PCR amplification of the gene coding for ribulose-1,5-bisphosphate carboxylase/oxygenase (rbcL) was successfully performed following all three fixation protocols. As expected the PCR product was restricted to the plastids of all cells. Reverse transcribed in situ PCR was performed on rbcL mRNA and in this case the PCR product was restricted to the plastids of the bundle sheath cells. This is the first report of in situ PCR on plant material and only the second report of in situ PCR with sub-cellular resolution. In situ PCR on rbcL may prove to be a valuable positive control for future in situ PCR studies on plant material.

AB - In situ PCR and reverse transcribed in situ PCR have been tested on leaves of sugar cane fixed in FAA, 4% PFA or 2% PFA+2.5% GA. In situ PCR amplification of the gene coding for ribulose-1,5-bisphosphate carboxylase/oxygenase (rbcL) was successfully performed following all three fixation protocols. As expected the PCR product was restricted to the plastids of all cells. Reverse transcribed in situ PCR was performed on rbcL mRNA and in this case the PCR product was restricted to the plastids of the bundle sheath cells. This is the first report of in situ PCR on plant material and only the second report of in situ PCR with sub-cellular resolution. In situ PCR on rbcL may prove to be a valuable positive control for future in situ PCR studies on plant material.

KW - c -plants

KW - in situ pcr

KW - pcr

KW - rbc l

KW - reversed transcribed in situ

UR - https://www.scopus.com/pages/publications/0030703198

U2 - 10.1006/anbo.1997.0502

DO - 10.1006/anbo.1997.0502

M3 - Journal article

SN - 0305-7364

VL - 80

SP - 697

EP - 700

JO - Annals of Botany

JF - Annals of Botany

IS - 5

ER -

In situPCR on Plant Material with Sub-cellular Resolution

Abstract

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