The Nucleus — Cytological Methods and Isolation for Biochemical Studies

  • J. F. Jackson
Part of the Modern Methods of Plant Analysis book series (MOLMETHPLANT, volume 1)

Abstract

Interest in the plant cell nucleus has heightened in recent times due to the rapid advances being made in the biochemistry of its DNA, RNA, and protein components. While the cytological observations on chromosome structure and behavior have held interest in the nucleus for some time, application of recombinant DNA technology, sequencing and hybridization techniques to chromosomal DNA have focussed attention on the nucleus even more. Studies on gene expression have also seen many advances recently and much has been learnt about the events of eukaryotic mRNA biogenesis (Nevins 1983). Recognition of promoters and sites of transcriptional initiation, poly(A) addition, RNA splicing, methylation and the process of nuclear-cytoplasmic transport have been investigated and all need nuclear preparations at some stage for further investigation in the eukaryotic organism. Possession of a well-defined nucleus is one of the criteria separating eukaryotes from prokaryotes, and it is the pathway of mRNA synthesis involving this RNA processing and transport out of the nucleus which sets the eukaryotes apart from the prokaryotes as much as the other cytologically recognized nuclear differences. There is then great interest in nuclear preparation for these and other biochemical studies. This chapter will deal in the main with the larger-scale preparation of nuclei from plant cells for these biochemical studies. The properties of the nucleus will be dealt with only in so far as it explains the various techniques exploiting those properties which are used in nuclear preparation. Particular attention will be given to different procedures adopted for nuclear preparations for different purposes. The development of efficient protoplast preparation methods from plant cells has had a great effect on organelle investigations in plants (see Wagner, this vol.) including nuclear studies.

Keywords

Starch Sedimentation Mannitol Cyclohexane Xylene 

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© Springer-Verlag Berlin Heidelberg 1985

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  • J. F. Jackson

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