. The Biological bulletin. Biology; Zoology; Marine biology. Figure 2. Northern blot hybridization on RNA extracted from the distal part and the rest of the foot of Mytihis gallopmvincialis. Ten mi- crograms of total RNA was electrophoresed on a agarose gel, trans- ferred onto a nylon membrane, and hybridized with ^-P-labeled cDNA encoding the whole coding region of Mgfp-1 and Mgfp-2. (A) Mgfp-1 sequence as a probe; (B) Mgfp-2 sequence as a probe. 1, the distal part of the foot; 2, the remainder of the foot. Arrowheads indicate the position of 18Sand28SrRNA. estimate the expression sites
. The Biological bulletin. Biology; Zoology; Marine biology. Figure 2. Northern blot hybridization on RNA extracted from the distal part and the rest of the foot of Mytihis gallopmvincialis. Ten mi- crograms of total RNA was electrophoresed on a agarose gel, trans- ferred onto a nylon membrane, and hybridized with ^-P-labeled cDNA encoding the whole coding region of Mgfp-1 and Mgfp-2. (A) Mgfp-1 sequence as a probe; (B) Mgfp-2 sequence as a probe. 1, the distal part of the foot; 2, the remainder of the foot. Arrowheads indicate the position of 18Sand28SrRNA. estimate the expression sites of the two genes in the foot, RNA was isolated separately from the distal end and from the remainder of the foot and was also analyzed by north- em blotting. The distal end of the foot consisted mainly of the phenol gland, but included a small fraction of the accessory gland, because there is a hairpin turn of the accessory gland interdigitated with the underlying phenol gland (Waite, 1992). As a result, Mgfp-1 mRNA was de- tected in both the distal end and the remainder of the foot, whereas Mgfp-2 mRNA was detected primarily in the distal end and was undetectable in the remainder of the foot. Moreover, the level of Mgfp-1 mRNA accu- mulation was higher in the remainder of the foot than in the distal end (Fig. 2). These results indicated that the Mgfp-1 and Mgfp-2 genes were expressed at diiferent sites in the foot. The length of Mgfp-1 cDNA was about kb (Inoue and Odo, 1994). This size was consistent with the position of the bands detected in Figures 1A and 2A. The length of Mgfp-2 cDNA was about kb (Inoue et al, 1995), and this size was also consistent with the position of the bands detected in Figures 1B and 2B. Mgfp-1 In situ hybridization The foot is an organ that has many functions. It con- tains various types of tissues and glands (Waite, 1992). To determine exact sites expressing the Mgfp-1 and Mgfjp- 2 genes in the foot, in situ hybridization was performed. The
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