|Budget Amount *help
¥2,500,000 (Direct Cost : ¥2,500,000)
Fiscal Year 1999 : ¥800,000 (Direct Cost : ¥800,000)
Fiscal Year 1998 : ¥1,700,000 (Direct Cost : ¥1,700,000)
The aim of this study is to establish the method of DNA flngerprin6g using PCR with arbitrary primers, which is then to be used for the future study on the social structure of the primitive, polygynous ant, Amblyopone sivlestrii. A. silvestrii is a small ant, whose single workers yielded c.400-700ng of genomic DNA. With this DNA as a template, the following synthetic oligomers were used as PCR primers designed to anchor 3' or 5' end of microsatellite loci: 1) Degenerate primers for CAP-PCR (3'-anchord primers), I.e., (CA)7 or (AC)7-DG or DA or DT or DC, and (GA)7 or (AG)7-HG or HA or HT or HC. 2) (GA)7ACTG, (GA)7ACTA, (GA)7ACTT, (GA)7ACTC, (AG)7CGTG, (AG)7CGTA, (AG)7CGTT, and (AG)7CGTC. 3) The same repeats as the former but anchored at 3' end with 2 arbitrary nucleotide residues. 4) 6 repeats of GA or GT anchored at 5' end with 3 to 5 arbitrary nucleotide residues. 5) Repeats without anchor sequence such as (CAG)5, (TAG)5, (GATA)4 and (GCGT)4.
Among these primers, one primer of the above category 2 has successfully produced polymorphism between individuals of A. silvestrii. Another primer of the same category also detected individual difference only when combined with RE-digestion. Although these primers are useful for the analysis of kin-structure, especially centered on polygyny, of this ant, yet we need some further primers.