South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Key Laboratory of South China Sea Fishery Resources Exploitation and Utilization, Ministry of Agriculture, Guangzhou, China.
Key Laboratory of Aquatic Genomics, Ministry of Agriculture, Beijing, China.
PLoS One. 2018 May 24;13(5):e0198036. doi: 10.1371/journal.pone.0198036. eCollection 2018.
Chk1 is a cell-cycle regulator. Chk1 has been identified in organisms ranging from yeast to humans, but few researchers have studied Chk1 in shrimps. We cloned Chk1 from the black tiger shrimp (Penaeus monodon). The full-length cDNA sequence of PmChk1 had 3,334 base pairs (bp), with an open reading frame of 1,455 bp. The complete genomic sequence of PmChk1 (11,081 bp) contained 10 exons separated by nine introns. qRT-PCR showed that PmChk1 was highly expressed in the ovaries and gills of P. monodon. The lowest PmChk1 expression was noted in stage III of ovarian development in P. monodon. PmChk1 expression decreased significantly after injection of 5-hydroxytryptamine and eyestalk ablation in P. monodon ovaries. RNA interference experiments were undertaken to examine the expression of PmChk1, PmCDC2, and PmCyclin B. PmChk1 knockdown in the ovaries and hepatopancreas by dsRNA-Chk1 was successful. The localization and level of PmChk1 expression in the hepatopancreas was studied using in situ hybridization, which showed that data were in accordance with those of qRT-PCR. The Gonadosomatic Index of P. monodon after dsRNA-Chk1 injection was significantly higher than that after injection of dsRNA-GFP or phosphate-buffered saline. These data suggest that PmChk1 may have important roles in the ovarian maturation of P. monodon.
Chk1 是一种细胞周期调控因子。Chk1 已在从酵母到人类等生物中被发现，但很少有研究人员在虾类中研究 Chk1。我们从黑虎虾（Penaeus monodon）中克隆了 Chk1。PmChk1 的全长 cDNA 序列有 3334 个碱基对（bp），开放阅读框为 1455bp。PmChk1 的完整基因组序列（11081bp）包含 10 个外显子和 9 个内含子。qRT-PCR 显示，PmChk1 在黑虎虾的卵巢和鳃中高度表达。在黑虎虾卵巢发育的 III 期，PmChk1 的表达最低。在注射 5-羟色胺和切除眼柄后，P. monodon 卵巢中 PmChk1 的表达显著下降。进行了 RNA 干扰实验以检查 PmChk1、PmCDC2 和 PmCyclin B 的表达。通过 dsRNA-Chk1 成功地在卵巢和肝胰腺中敲低了 PmChk1。使用原位杂交研究了 PmChk1 在肝胰腺中的定位和表达水平，结果表明数据与 qRT-PCR 的结果一致。dsRNA-Chk1 注射后黑虎虾的性腺指数显著高于 dsRNA-GFP 或磷酸盐缓冲液注射后的性腺指数。这些数据表明，PmChk1 可能在黑虎虾的卵巢成熟过程中发挥重要作用。
