Postendocytic Provitellin Processing in the Growing Oocyte of the Short Horned Grasshopper, Oxyajaponicajaponica (Orthoptera: Acrididae)

Abstract

Polyclonal antibodies made against 86 kDa (86 k), 80 kDa (80 k) and 54 kDa (54 k) vitellins of Oxya japonica japonica are used for Western blotting. Anti‐80k vitellin antibody is cross‐reacted with a 95 kDa (95 k) vitellin. While 95 k vitellin is present both in the female hemolymph and in the oocyte, 80 k vitellin is detected only in the oocyte and the laid egg. In the growing oocytes, as 95 k vitellin is faded out gradually, 80 k vitellin is accumulated increasingly, indicating postendocytic processing of 95 k vitellin brings 80 k vitellin. Further conforming the hypothesis, partial digestion of 95 k vitellin with pepsin and α‐chymotrypsin makes several protein bands of molecular weight around 80 kDa. Thus, the 95k vitellin may have a cleavage site (s) to produce 80 k vitellin which forms fairly stable tertiary structure. In the reduced condition (20 mM glutathion), both 95 k and 80 k vitellins were digested throughly by endogenous proteinase at pH 4. Both 86 k and 54 k vitellins, respectively, show no apparent molecular weight changes in the growing oocyte and in the hemolymph.

Isolation and expression analysis of a homolog of the 14–3–3 epsilon gene in the diamondback moth, Plutella xylostella

“…The gel slices containing overexpressed proteins was homogenized and mixed with adjuvant. The mixture was injected into a rabbit to generate anti-Px14-3-3e IgG (called Px14-3-3e-Ab1) (Cho and Park, 2004).…”

cDNA cloning and induction of tyrosine hydroxylase gene from the diamondback moth, Plutella xylostella

“…The gel slices containing over-expressed proteins were homogenized and mixed with an adjuvant. The mixture was injected into a rabbit to generate anti-PxTH IgG (called PxTH Ab-2) (Cho and Park, 2004). We also ordered a rabbit antibody, called PxTH-Ab2, against a fragment of the PxTH protein (amino acids 8 through 27; QKNREMFAIKKSYSIENGYP) from Peptron Corp., Korea.…”