Mutagenesis testing using transgenic non-human animals carrying test DNA sequences

A mutagenesis testing method comprising:

• (a) exposing a transgenic mouse to a test agent, said transgenic mouse comprising somatic and germ cells containing a test DNA sequence recoverable from said cells via enzymatic excision sites, the expression of said sequence being capable of detection in bacteria by bioassay, wherein said bioassay selects for cells containing mutations in said test DNA sequence, wherein said test sequence is selected from the group consisting of LacZ, LacZ.alpha., LacI and LacI.sup.Q ;
• (b) recovering a sample of said test DNA sequence from said exposed mouse, wherein said recovery is mediated throuqh enzymatic excision of a type functional for lambda cos sites or lambda ZAP excision sites;
• (c) treating said recovered sample of step (b) with a restriction system deficient bacteriophage packaging extract to package said recovered test DNA sequence of step (b);
• (d) infecting a phage-free, prolysogenic strain of E. coli with said packaged test DNA sequence;
• and
• (e) determining by bioassay of said infected E. coli of step (d) the frequency of mutation of said test DNA sequence.

A mutagenesis testing method comprising:

• (a) exposing a transgenic mouse to a test agent, said transgenic mouse comprising somatic and germ cells containing a test DNA sequence contained within a system permitting enzymatic excision and circularization of the test DNA sequence, wherein said test sequence is selected from the group consisting of LacZ, LacZ.alpha., LacI and LacI.sup.Q ;
• (b) recovering a sample of said test DNA sequence from said exposed mouse, wherein said recovery is mediated through enzymatic excision of a type functional for lambda cos sites or lambda ZAP excision sites;
• and
• (c) determining by bioassay in a prolysogenic E. coli that is restriction system deficient the frequency of mutation of said test DNA sequence in said recovered sample, wherein said bioassay selects for cells containing mutations in said test DNA sequence.

A mutagenesis testing method comprising:

• (a) exposing a transgenic mouse to a test agent, said transgenic mouse comprising somatic and germ cells containing a test DNA sequence contained within a system permitting enzymatic excision and circularization of the test DNA sequence, wherein said test sequence is selected from the group consisting of LacZ, LacZ.alpha., LacI and LacI.sup.Q ;
• (b) recovering a sample of said target DNA sequence within said system from said exposed mouse, wherein said recovery is mediated throuqh enzymatic excision of a type functional for lambda cos sites or lambda ZAP excision sites;
• (c) excising and circularizing the test DNA sequence in said recovered sample using said system;
• and
• d) determining by bioassay in a phage-free prolysogenic strain of E. coli that is restriction system deficient the frequency of mutation of said test DNA sequence in said circularized sample, wherein said bioassay selects for cells containing mutations in said test DNA sequence.

A mutagenesis testing method comprising:

• (a) exposing a transgenic mouse to a test agent, said transgenic mouse comprising somatic and germ cells containing a test DNA sequence flanked on both sides by one of two halves of an f1 bacteriophage origin of replication contained by a bacteriophage origin of replication contained by a bacteriophage derivative genome, wherein said test sequence is selected from the group consisting of LacZ, LacZ.alpha., LacI and LacI.sup.Q ;
• (b) rescuing with f1 helper phage proteins a sample of said test DNA sequence from said exposed mouse;
• and
• (c) determining the frequency of mutation of said test DNA sequence in said rescued sample by bioassay in a prolysogenic E. coli that is restriction system deficient, wherein said bioassay selects for cells containing mutations in said test DNA sequence.

A mutagenesis testing method comprising:

• (a) exposing a transgenic mouse to a test agent, said transgenic mouse comprising somatic and germ cells containing a test DNA sequence flanked on both sides by one of two halves of an f1 bacteriophage origin of replication contained by a bacteriophage origin of replication contained by a bacteriophage derivative genome, wherein said test sequence is selected from the group consisting of Lacz LacZ.alpha., LacI and LaCI.sup.Q ;
• (b) recovering a sample of said test DNA sequence from said exposed mouse;
• (c) excising a sample of the recovered test DNA sequence with f1 helper phage proteins;
• (d) determining the frequency of mutation of said test DNA sequence in said excised sample by bioassay in a prolysogenic E. coli that is restriction system deficient, wherein said bioassay selects for cells containing mutations in said test DNA sequence.