Yang Fan, Xu Hua-Teng, Dai Zhong-Min, Yang Wei-Jun
College of Life Sciences, Zhejiang University, 232 Wensan Road, Hangzhou, Zhejiang, 310012 PR China.
Comp Biochem Physiol B Biochem Mol Biol. 2005 Dec;142(4):456-64. doi: 10.1016/j.cbpb.2005.09.011. Epub 2005 Oct 27.
As a precursor of the major yolk protein vitellin (Vn), vitellogenin (Vg) has been studied in crustacean for decades, mainly in shrimp. However, little is known about the Vg molecule in crabs. In the present study, we report a cDNA encoding Vg in the marine crab Portunus trituberculatus, an important commercial species in aquaculture. The cDNA sequence is 7,846 bp in length, containing a 7,680 bp open reading frame, that encodes 2,560 amino acid residues. The deduced amino acid sequence showed 32-75% identity with other known crustacean Vgs. Seven potential cleavage sites (RXXR) were identified in the deduced Vg precursor. SDS-PAGE and Western blotting analysis demonstrated that Vns in the ovary of the mature crab consisted of three major polypeptides (102, 100 and 85 kDa). Northern blotting analysis revealed that Vg was mainly expressed in hepatopancreas in vitellogensis females. A Vg transcript was also identified in the ovary of the crab by reverse transcription-polymerase chain reaction analysis.
作为主要卵黄蛋白卵黄磷蛋白(Vn)的前体,卵黄原蛋白(Vg)已在甲壳类动物中研究了数十年,主要是在虾类中。然而,关于蟹类中的Vg分子却知之甚少。在本研究中,我们报道了一种在海洋蟹类三疣梭子蟹(Portunus trituberculatus)中编码Vg的cDNA,三疣梭子蟹是水产养殖中的一种重要商业品种。该cDNA序列长度为7846 bp,包含一个7680 bp的开放阅读框,编码2560个氨基酸残基。推导的氨基酸序列与其他已知甲壳类动物的Vg具有32%-75%的同一性。在推导的Vg前体中鉴定出七个潜在的切割位点(RXXR)。SDS-PAGE和蛋白质印迹分析表明,成熟蟹卵巢中的Vn由三种主要多肽(102、100和85 kDa)组成。Northern印迹分析显示,Vg主要在卵黄发生期雌性的肝胰腺中表达。通过逆转录-聚合酶链反应分析,在蟹的卵巢中也鉴定出了一个Vg转录本。