Cetus Corporation

Chander P Bahl

Jack H Nunberg

Recombinant plasmids are described that carry the structural gene for human proinsulin in expressible form. The human proinsulin structural gene is fused to a functional transport sequence. Host organisms transformed by the recombinant plasmids express a preproinsulin in vivo, and is transported across the host organism's cell wall. Following its recovery, in vitro methods are used to convert the human proinsulin to human insulin.

Plasmid for producing human insulin

A method is described for constructing double-stranded DNA using messenger RNA templates. DNA strands complementary to the templates are formed using a non-specific 3' hydroxy terminus primer. After separation, the DNA single strands are disabled from self priming by blocking the free 3' hydroxy. Primer segments are then provided complementary to a portion of the amino terminus-coding end of the desired single strands. These primer segments are then extended with a suitable polymerase to complete the complementary strands.

Method for using an amino-terminus DNA sequence to synthesize a specific

A method for altering a single nucleotide at a predetermined position in a gene. The method involves the isolation of a single strand gene fragment extending up to the position before the nucleotide to be altered. A ribonucleotide or a protected deoxyribonucleotide corresponding to the desired altered nucleotide is attached at the end of this fragment. The fragment is then annealed to a complementary template which extends beyond the end of the fragment. The fragment is then extended complementary to the remainder of the template. The resulting partially mismatched double-stranded DNA is used to produce a pure DNA gene containing an altered deoxyribonucleotide at the single desired position.

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El Cerrito, CA, United States of America

Chander P Bahl