Animal Virus Genetics by Bernard N. Fields and Rudolf Jaenisch (Eds.)

By Bernard N. Fields and Rudolf Jaenisch (Eds.)

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Experiments are in progress to further characterize these clones. Of the 7 bands not represented among our clones, at least two must represent the ends of the genome and cannot be cloned using the present method because only one terminus of the viral fragment will have an Eco RI site. The HCMV Eco RI fragments which correspond to the ends of the genome are currently being identified, and will be cloned after gel purification of those bands and the attachment of Eco RI linkers. More than two end fragments may exist if the HCMV genome is organized in a manner similar to that of Herpes Simplex, with long and short regions which occur in four different arrangements (see 19 for review).

25 molar fragments in certain restriction enzyme digests of HSV DNA, especially those of the HSV-KMP) strain with HindIII and Bglll (12,18). 25M pattern. 0M species. An examination of the Hindlll cleavage pattern reveals at least eight submolar species with varying molarities. 5M. For convenience, during the initial stages of the mapping analysis, these submolar species are referred to as ml, m2, m3, etc. in each digest and the molar fragments are referred to by letters A, B, C, etc. in order of decreasing size.

High molecular weight DNA from either uninfected human cells or human 24 JOYCE C. TAMASHIRO AND DEBORAH H. EcoRt+BAP Human Fibroblast mm <3Ξ=> CMV Virions Purification of CMV DNA / Eco R, cuts CMV DNA into many fragments in of different sizes. Annealing and Ligation o oOoo Mixing of recombinant plasmids with E. coli cells in high Ca + + <**ol I^PQI k>ol |i*a- oj Selection of clones of E. coli each bearing a recombinant plasmid (Tet R; Chlors) FIGURE 1. Strategy for the cloning of human cytomegalovirus DNA.

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