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Nucleic Acids Research, 1978, Vol. 5, No. 9 3275-3294
© 1978

Articles

Cloning of chicken lysozyme structural gene sequences synthesized in vitro

Albrecht E. Sippel, Hartmut Land, Werner Lindenmaier, M.Chi Nguyen-Huu, Wurtz Tilmann, Kenneth N. Timmis, Klaus Giesecke and Günther Schütz

Max-Planck-lnstitut für Molekulare Genetik Ihnestrasse 63-73, D-1000 Berlin 33, GFR

Received July 14, 1978. Double-stranded chicken lysozyme cDNA was synthesized from an oviduct' mRNA fraction enriched for lysozyme mRNA. The ds-cDNA was inserted into the BamHI site of plasmid pBR322 using chemically synthesized DNA linker molecules containing the BamHI restriction endonuclease cleavage site. After bacterial transformation, colonies carrying lysozyme DNA were identified by hybridization with highly purified lysozyme cDNA. The 555 base pairs long cloned DNA fragment of one recombinant plasmid was isolated and characterized by restriction endonuclease digestion. The DNA sequence of selected parts of the inserted DNA is as predicted from the amino acid sequence of prelysozyme. The sequence data allows the unambiguous location of the coding region within lysozyme mRNA.


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