Process for obtaining DNA, RNA, peptides, polypeptides, or proteins, by recombinant DNA technique

A process for the production of a host cell capable of producing a transcription product or a translation product comprising an expression vector, wherein said expression vector comprises at least one stochastic sequence of polynucleotides comprising the steps of:

• producing a library of expression vectors capable of producing a transcription product or a translation product, said expression vectors comprising at least one stochastic sequence of polynucleotides, said expression vectors being produced by the following steps:
• linearizing an expression vector;
• reacting said linearized expression vector with terminal transferase enzyme in the presence of desired ratios of deoxynucleotide-triphosphates of guanine, cytosine, thymidine, and adenine to form a stochastic polynucleotide sequence at each 3' extremity of said linearized vector;
• hybridizing said stochastic polynucleotide sequence at a 3' extremity of said linearized expression vector;
• synthesizing a second strand from said 3' ends of said hybridized expression vector by incubating with polymerase;
• and
• transforming a host cell with said expression vector.

A process for the production of a host cell capable of producing a transcription product or a translation product comprising an expression vector, wherein said expression vector comprises at least one stochastic sequence of polynucleotides comprising the steps of:

• producing a library of expression vectors capable of producing a transcription product or a translation product, said expression vectors comprising at least one stochastic sequence of polynucleotides;
• transforming a host cell with said expression vector;
• culturing said transformed host cell;
• screening and/or selecting said transformed host cell;
• and
• isolating said selected or screened host cell.

A method of producing a diverse population of host cells comprising:

• (a) synthesizing a diverse population of stochastically generated polynucleotide sequences comprising greater than about 1.times.10.sup.5 different polynucleotide sequences, said method consisting of stochastic copolymerization of double stranded oligonucleotides, copolymerizaton of the four kinds of nucleotide triphosphates consisting of adenine, cytosine, guaninie and thymine, and chemical synthesis, and
• (b) inserting said diverse population of stochastically generated polynucleotide sequences into a population of vectors to form a diverse population of vectors containing stochastically generated polynucleotide sequences;
• and
• (c) inserting said diverse population of vectors into host cells.

A method of producing a diverse populations of host cells, comprising:

• (a) obtaining one or more diverse populations of vectors containing diverse sequences of double stranded polynucleotides;
• (b) digesting the one or more diverse populations of vectors with a restriction enzyme, and
• (c) stochastically copolymerizing the one or more diverse populations of double stranded polynucleotides so as to produce a new population of greater than about 1.times.10.sup.5 different polynucleotide sequences;
• and
• (d) inserting said new population of polynucleotides into host cells.