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Wu X.,Ocean University of China | Wu X.,Key Laboratory of Marine Genetics and Breeding | Wang Z.,Ocean University of China | Wang Z.,Key Laboratory of Marine Genetics and Breeding | And 10 more authors.
Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology | Year: 2013

Vasa is a DEAD box helicase and has shown essential functions during gametogenesis and embryogenesis. In most species, research revealed a specific expression of vasa gene in the germ cells. Thus, vasa has become the candidate gene in identifying germ cells. In this study, the vasa gene was isolated from gonads of Japanese flounder ( Paralichthys olivaceus). In the 11.4. kb genomic sequence, 23 exons were identified besides 5' and 3' flanking regions. The promoter region contained several putative TF binding sites which may have the function of regulating vasa expression. Quantitative real-time PCR analysis showed that vasa gene expression was restricted to adult gonads, with a higher level in the ovary. Development expression profiling revealed a maternal deposit and constant embryonic expression at early stages, but the relative mRNA amount decreased after gastrula. Nine other PoVasa transcripts were detected and their expression in gonads and during early development was not all the same, implying potential different functions during gametogenesis or early embryonic development. These results together confirmed the feasibility of using vasa as a marker of germ cells and that vasa gene had an important role in spermatogenesis and oogenesis. Furthermore, our study laid the groundwork for identifying fish primordial germ cells (PGCs) and investigating germ cell biology. © 2013 Elsevier Inc.

Han Q.,Key Laboratory of Marine Genetics and Breeding | Dong D.,Key Laboratory of Marine Genetics and Breeding | Zhang X.,Key Laboratory of Marine Genetics and Breeding | Liang C.,Key Laboratory of Marine Genetics and Breeding | And 3 more authors.
Crustaceana | Year: 2015

In this study, both liposome- And retrovirus-mediated gene transfer methods were examined for their potential to transfer and express two retroviral vectors containing the mouse c-Myc or the green fluorescent protein (GFP) gene into the primary lymphoid cell cultures (OKA) derived from "Oka" organs (= organs of the lymphoid system) of the greasyback shrimp Metapenaeus ensis (De Haan, 1844). It was found that the c-Myc gene could be delivered into OKA cells by the liposomemediated method, but the introduced c-Myc gene could not be effectively transcribed into mRNA. In contrast, the pantropic retrovirus-mediated method failed to introduce the c-Myc gene into OKA cells, and GFP was not detected in the transformed cells, either. This work inferred two problems for the use of the two above-mentioned gene transfer methods in the non-dividing OKA cells: (1) the viral promoter of long terminal repeats (LTRs) had low activity in shrimp cells; (2) the pantropic retrovirus-mediated gene transfer system had a low tropism to shrimp lymphoid cells. © Koninklijke Brill NV, Leiden, 2015.

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