Characterization, expression and silencing by RNAi of p53 from Penaeus monodon.
The tumor suppressor p53 is a sequence-specific transcription factor, whose target genes can regulate genomic stability, the cellular response to DNA damage and cell-cycle progression. In the present study, the full-length complementary DNA (cDNA) sequence of p53 gene from Penaeus monodon (Pmp53) was cloned by the technology of rapid amplification of cDNA ends (RACE). The cDNA of Pmp53 was 2239 bp, encoding a protein of 450 amino acids with calculated molecular weight of 50.62 kDa. The temporal expression of Pmp53 in different tissues (ovary, heart, intestine, brain, muscles, stomach and gills) and different developmental stages of ovary was investigated by real-time quantitative PCR (RT-qPCR). The lowest expression level of Pmp53 was observed in the stomach, while the highest expression level was detected in the brain. During the ovary development stages, the expression level of Pmp53 reached the peak at stage III. RNA interference (RNAi) and serotonin (5-hydroxytryptamine, 5-HT) injection experiments were conducted to study the expression profile of Pmp53 and PmCDK2 (cyclin-dependent kinase 2, CDK2). Knocked down of Pmp53 by dsRNA-p53 was sequence-specific and successful. Expression levels of Pmp53 and PmCDK2 in ovary of P. monodon were significantly increased at 12-96 h post 5-HT injection. These results indicate that Pmp53 may be involved in the regulation of ovarian development of P. monodon.
South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China.|College of Aqua-life Science and Technology, Shanghai Ocean University, Shanghai, China.
Wenting
W
Dai
South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China. qiu902@yahoo.com.|Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture, Guangzhou, China. qiu902@yahoo.com.
Lihua
L
Qiu
South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China.|Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture, Guangzhou, China.
Chao
C
Zhao
South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China.|Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture, Guangzhou, China.
Mingjun
M
Fu
South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China.|Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture, Guangzhou, China.
Zhenhua
Z
Ma
South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China.|Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture, Guangzhou, China.
Falin
F
Zhou
South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China.|Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture, Guangzhou, China.|Tropical Aquaculture Research and Development Center of South China Sea Fisheries Research Institute, Sanya, China.
Qibin
Q
Yang
D000595:Amino Acid Sequence; D000818:Animals; D060829:Arthropod Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D005260:Female; D015870:Gene Expression; D043143:Hepatopancreas / Q000378:metabolism; D008297:Male; D009928:Organ Specificity; D010053:Ovary / Q000254:growth & development / Q000378:metabolism; D033561:Penaeidae / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D010802:Phylogeny; D011499:Protein Processing, Post-Translational; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000235:genetics* / Q000378:metabolism
D060829:Arthropod Proteins; D012333:RNA, Messenger; D016159:Tumor Suppressor Protein p53
Gene cloning; Ovarian development; Penaeus monodon; Pmp53; RNAi; RT-qPCR