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- W1020346317 abstract "Publisher Summary This chapter describes methods for the direct genetic engineering of the cyanobacterial chromosome and techniques for the analysis of DNA and RNA from the resulting transformants. The DNA in lane B described in the chapter is a by-product of the RNA isolation procedure. The genetic engineering of the cyanobacterial chromosome includes the techniques described in the chapter. The chapter focuses on methods that employ wholly homologous transforming DNA or heterologous sequences, which are flanked by cyanobacterial DNA on either side. The analysis of DNA from transformants is necessary to determine the chromosomal structure following recombination. The recombination events illustrated in the chapter are those that are usually seen, but aberrant events have been observed when the cloned region homologous to the chromosome is small. These events include the deletion of a chromosomal segment and integration at a related locus in the case of a gene family. Southern hybridization can determine whether the expected event has in fact, occurred. It is also important to analyze RNA from transformants, which have been engineered by these methods. A homologous recombination event, which replaces a wild-type gene with another functional allele, should result in the production of a full-length mitochondrial RNA (mRNA), which initiates and terminates properly. The chapter also mentions the instructions of the procedure that describe the preparation of glass fines used in the cyanobacterial total DNA miniprep procedure, which was adapted from the method of Vogelstein and Gillespie." @default.
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- W1020346317 date "1987-01-01" @default.
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- W1020346317 title "[12] Genetic engineering of the cyanobacterial chromosome" @default.
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- W1020346317 doi "https://doi.org/10.1016/0076-6879(87)53055-5" @default.
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