[Expression of a gene fragment from the alpha-subunit of the acetylcholine receptor from Torpedo californica in Saccharomyces cerevisiae].

A fragment of the AChR alpha-subunit gene from Torpedo californica (about 800 bp) was joined in frame with a synthetic duplex, coding for a changed leader peptide of the Kluyveromyces lactis killer toxin under the control of the reconstructed glyceraldehyde-3-phosphate dehydrogenase gene promoter in the pUC8 vector. Translation termination codons in all frames were inserted as a part of a self-complementary oligodeoxynucleotide to the Eco47III site (807 bp from the start of the gene). Vector pJDAch was constructed by cloning the expression cassette between the BamHI and HindIII sites in the multicopy yeast plasmid pJDB207.

[Chemical-enzymatic synthesis and cloning in Escherichia coli of a gene coding for human granulocyte-colony stimulating factor].

Two artificial genes, encoding two forms of human granulocyte colony stimulating factor as products of a normal and an alternative splicing, have been by a chemical-enzymatic way synthesized and cloned in Escherichia coli. The genes are supplied with recognition sites of restriction endonucleases to facilitate the further cassette mutagenesis.

[A new approach to DNA synthesis from synthetic oligonucleotides. Synthesis of DNA coding for the repeated antigenic determinant of foot and mouth disease virus].

A rapid method for assembly of DNA from synthetic oligodeoxynucleotides has been developed which involves separate ligation of top- and bottom-strand oligonucleotides followed by filling in 3'-ends of the duplex formed, blunt end cloning into a specialized vector pBBV, and recovery of the synthetic DNA from the recombinant plasmid by means of restriction nuclease BbvII. The method allows for many oligonucleotides to be ligated at once, with no intermediates being isolated, and any DNA to be recovered on cloning, no matter what the sequences of its termini are. Ten oligodeoxynucleotides (I)-(X) have been chemically synthesised and used to prepare, by this method, a 60-membered duplex with complementary tetranucleotide 5'-protrusions (DNA I) which comprises the cDNA sequence 3397-3456 of foot and mouth disease virus (FMDV) strain O1K.