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Nucleic Acids Research, 1986, Vol. 14, No. 15 6159-6168
© 1986

Articles

Simultaneous synthesis of human-, mouse- and chimeric epidermal growth factor genes via ‘hybrid gene synthesis’ approach

Wing L. Sung*, Diana M. Zahab, Fei-L. Yao, Ray Wu+ and Saran A. Narang

Division of Biological Sciences, National Research Council of Canada Ottawa, Ontario K1A 0R6 +Section of Biochemistry, Molecular and Cell Biology, Cornell University Ithaca, NY 14853, USA

*To whom all the correspondence should be addressed.

Received March 3, 1986. Revised June 30, 1986. Accepted June 30, 1986.

Simultaneous synthesis of two DNA duplexes encoding human and mouse epidermal growth factors (EGF) was accomplished in a single step. A 174 b.p. DNA heteroduplex, with 16 single and double base pair mismatches, was designed. One strand encoded the human EGF, and the opposite strand indirectly encoded the mouse EGF. The heteroduplex DNA was synthesized by ligation of seven overlapping oligodeoxyribonucleotides with a linearized plasmid. After transformation in E. coli HB101 (recAl3) , the resulting heteroduplex plasmid served as the template in plasmid replication. Two different plasmid progenies bearing either the human or mouse EGF-coding sequence were identified by colony hybridization using the appropriate probes. However, in E. coli JM103, the same process yielded plasmid progenies encoding different chimeric EGF molecules, presumably due to crossover of human and mouse EGF gene sequences.


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