Allele frequencies and their variances were calculated from genotype frequencies observed in zymograms
using the BIOSYS-1 package (Swofford and Selander 1989).
The study of inhibitors both in vitro and through the zymogram
in adults of M.
Caption: Figure 1: SDS-PAGE with corresponding zymogram
of AmyMH expressed in Brevibacillus choshinensis SP3.
The SDS-PAGE analysis of anion-exchange fractions revealed a single protein band of approximate 25 kDa as shown in Figure 2A, lane 4, that corresponds to the only active band seen on casein zymogram
, presenting the same molecular weight (Figure 2B).
of both testicular samples revealed only single band at identical position.
A strip of the gel was cut and bands were identified on zymogram
For the protease zymogram
, the proteins were first separated in nondenaturing conditions on IEF (3,0-9,0) Phastgels.
Apart from the inability of the zymogram
technique to detect all variation (allozyme bands effectively are electromorphs, that is, collections of similarly charged enzyme molecules that differ in their amino-acid sequence at one site at least; King and Ohta 1975), a given Taraxacum genotype may comprise a heterogeneous collection of individuals sharing an identical three-locus electrophoretic profile.
For activity staining (Zymogram
) of xylanase by SDS-Xylan-PAGE, SDS was removed by washing the gel at room temperature in solution-A (50 mM [Na.sub.2]HP[O.sub.4], 50 mM Na[H.sub.2]P[O.sub.4] (pH 7.2), isopropanol) for ne hr and solution-B (50 mM [Na.sub.2]HP[O.sub.4], 50 mM Na[H.sub.2]P[O.sub.4] (pH 7.2)) for one hr, respectively.
differences among the five species of the abalone from the coasts of Japan.
Cell culture media, antibiotics, and 10% Zymogram
gel were purchased from Life Technologies (Rockville, MD, USA).
Additionally, we have improved the protocols of electrophoresis, traditionary chromosomal preparation and exogenous gene transfection by trying a local apparatus, different buffers, colchicine and hypotonic time, plasmid DNA incubation time and lipofectamine combination, thus make these techniques feasible under our conditions and greatly improve isoenzyme zymogram
, karyotype quality and exogenous gene transfection efficiency.