Environmental and Molecular Mutagenesis - Legacy Tobacco ...
Environmental and Molecular Mutagenesis - Legacy Tobacco ...
Environmental and Molecular Mutagenesis - Legacy Tobacco ...
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122 1989 EMS Abstracts<br />
Notes HPRT gene, are used to amplify the first <strong>and</strong> second str<strong>and</strong> cDNA 5 x 107 fold during<br />
30 cycles of polymerase chain reaction (PCR) . The product (2 to 10 ng) is then<br />
purified by ultrafiltration, diluted 1 :1000, <strong>and</strong> subjected to an additional 30<br />
cycles of PCR, using two 20-mer primers located just interior to the first set .<br />
The product (-10 ug) is then sequenced directly using three end-labeled sequencing<br />
primers <strong>and</strong> Sequenase . With this system, we have sequenced 26 BPDE-induced mutants<br />
<strong>and</strong> found that 24/26 involved base substitutions . 97% of these involved G•C,<br />
predominantly G-C--*T•A, distributed throughout the 9 exons but with many located<br />
in exon 3 . (Supported by NCI Grant CA21253 .)<br />
http://legacy.library.ucsf.edu/tid/clb93d00/pdf<br />
I<br />
350<br />
ANEUPLOIDY FREQUENCIES IN MOUSE METAPHASE II OOCYTES AND FIRST-CLEAVAGE ZYGOTES<br />
FOLLOWING ORAL DOSAGES OF COLCHICINE . John B . Mailheal, Zhi Ping Yuanl, <strong>and</strong> M .J .<br />
Aardema2 . lDepartment of Obstetrics <strong>and</strong> Gynecology, L .S .U . Medical Center,<br />
Shreveport, LA 71130 ; 2The Proctor <strong>and</strong> Gamble Company, Cincinnati, OH 45247 .<br />
Certain chemicals can interact with cellular organelles responsible for orderly<br />
chromosome segregation <strong>and</strong> enhance the frequency of aneuploidy . Our objective was to<br />
determine the proportion of colchicine-induced aneuploid metaphase II (MII) oocytes<br />
that were transmitted to first-cleavage (lCl) zygotes . Female, CD-1 mice received 7 .5<br />
I .U . PMS <strong>and</strong> 5 .0 I .U . HCG was given 48 h later . Colchicine (2, 3, or 4 mg/kg) or<br />
sterile distilled water (solvent) was given by oral intubation iamnediately following<br />
HCG . MII oocytes were collected 16 h post HCG, whereas 1C1 zygotes were collected 16<br />
h post mating . The proportions (<strong>and</strong> percentages) of hyperploid MII oocytes were 3/216<br />
(1 .4), 81/512 (15 .8), 71/411 (17 .3), <strong>and</strong> 98/413 (23 .7) for control, 2 .0 mg/kg, 3 .0<br />
mg/kg, <strong>and</strong> 4 .0 mg/kg, respectively . Whereas, the proportions of hyperploid 1C1<br />
zygotes were 2/320 (0 .6), 23/364 (6 .3), 68/372 (18 .3), <strong>and</strong> 98/337 (29 .1) for control,<br />
2 .0 mg/kg, 3 .0 mg/kg, <strong>and</strong> 4 .0 mg/kg, respectively . The proportions of hyperploid<br />
cells differed (P0 .05) between MII oocytes <strong>and</strong> lCl zygotes in controls, <strong>and</strong> the 3 .0 mg/kg <strong>and</strong> 4 .0<br />
mg/kg groups . •the level of hyperploidy was greater (P
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