Institut Pasteur

Centre National De La Recherche Scientifique

Institut National De La Sante Et De La Recherche Medicale

Francis Galibert

Pierre Tiollais

Patrick Charnay

Yvon Cavaloc

The invention relates to a Happy Mapping type method for mapping a DNA molecule comprising a step enabling ad infinitum amplification of the DNA of each panel, consisting of the following: (i) a first amplification with the aid of a primer comprising 10-30 nucleotides defined at extremity 5&prime; thereof, and 5-10 random nucleotides at extremity 3&prime; thereof, in addition to (ii) a second amplification with the aid of a primer comprising at least the oligonucleotides defined at extremity 5&prime; of the primer used in stage i.

Method for mapping a DNA molecule comprising an ad infinitum amplification step

A process produces DNA corresponding to that of the DNA of the virus of B hepatitis. It comprises cloning in bacteria the latter DNA, previously repaired by means of the corresponding precursor nucleotides in the presence of a polymerase. Vectors contain the cloned DNA in their genomes. The cloned DNA is useful as a probe for detecting the presence of the virus of B hepatitis in biological samples, particularly blood or plasma. Its expression in bacteria provides a hybrid protein containing a protein fragment having vaccinating properties against hepatitis B. Nucleic acid of reduced size and a vector containing the nucleotide sequence of which DNA codes an immunogenic peptide sequence capable of inducing the generation of antibodies to the virus of viral hepatitis B. It comprises totally or partly the sequence of nucleotides represented in FIG. 9A. Application to the production by cloning in a bacterium of an immunogenic protein immunizing against hepatitis B, or application to the obtention of probes for the diagnosis of the presence of Dane particles in the serum.

Nucleotide sequence comprising the genome of hepatitis B virus,

Nucleotidic sequence coding the surface antigen of the hepatitis B

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Saint-Gratien, France

Francis Galibert