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Nucleic Acids Research, 1983, Vol. 11, No. 1 159-174
© 1983

MOLECULAR BIOLOGY

Methylation of the T-DNA in Agrobacterium tumefaciens and In several crown gall tumors

Stanton B.Gelvin, Susan J. Karcher and Victor J. DiRita

Department of Biological Sciences, Purdue University West Lafayette, IN 47907, USA

Received July 23, 1982. Accepted October 26, 1982.

Methylation of the T-DNA in Aqrobacterium tumefaciens and in four octopine-type (A6S/2, E9, 15955/1, 15955/01) and one nopaline-type (HT37#15) crown gall tumors was investigated using the isoschizomeric restriction endo-nucleases Msp I and Hpa II. T-DNA in the octop1ne-type Ti-plasmid pTiB6806 was not methylated at the sequence 5 ‘CCGG3’ in Agrobacterium. With two possible exceptions, neither was the T-DNA of the nopaline-type T1-plasm1d pTiT37 methylated in the bacterium. In all tumor lines investigated, at least one copy of the T-DNA was not methylated. DNA methylation was not detected in the lines A6S/2, 15955/1, HT37#15, and the TL region of E9. DNA methylation of some copies of TR in the E9 tumor line, and possibly in the 15955/01 line, was detected. Tne methylation of some copies of TR in the E9 line may indicate that not all copies of TR are transcribed in this tumor.


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