ABSTRACT: Identification of testis-relevant genes using in silico analysis from testes ESTs and cDNA microarray in the black tiger shrimp (Penaeus monodon)
Project description:The study aimed to determine effect of polychaetes as a shrimp feed on male reproductive maturation at transcriptional level through a cDNA microarray in the black tiger shrimp (Penaeus monodon). Thus, the experiment was to compare transcriptomic profiles of two different parts of reproductive organs, namely testes (TT) and vas deferens (VD), of domesticated 17-month-old between two different feeds, namely commercial pellet and polychaetes after feeding for one month. Differentially expressed genes were identified through the microarray analysis, and the microarray results were confirmed by real-time PCR. Selected genes were further characterized.
Project description:Comparisons of gene expression profiles from testis of wild and domesticated male brooders were made through a cDNA microarray in the black tiger shrimp (Penaeus monodon). Differentially expressed genes were identified through the microarray analysis, and the microarray results were confirmed by real-time PCR. Selected genes were further characterized.
Project description:Comparisons of gene expression profiles between testis and ovary of juvenile and wild brooders were made through a cDNA microarray in the black tiger shrimp (Penaeus monodon). Differentially expressed genes were identified through the microarray analysis, and the microarray results were confirmed by real-time PCR. Selected genes were further characterized.
Project description:Comparisons of gene expression profiles between ovaries of before (day 0) and after eyestalk ablation (days 1, 4 and 7) of domesticated 14-month-old black tiger shrimp (Penaeus monodon) were made using a cDNA microarray. Differentially expressed genes were identified through the microarray analysis, and the microarray results were confirmed by real-time PCR. Selected genes were further characterized.
Project description:Comparisons of gene expression profiles between testis and ovary of juvenile and wild brooders were made through a cDNA microarray in the black tiger shrimp (Penaeus monodon). Differentially expressed genes were identified through the microarray analysis, and the microarray results were confirmed by real-time PCR. Selected genes were further characterized. A cDNA microarray was constructed from the EST libraries of P. monodon, consisting of 5,568 features. RNA samples were extracted from both testes and ovaries of pooled juveniles (4 month-old, pooled from n=114 ), individual juvenile and broodstocks from West sides of Thailand. The RNA from ovaries were labeled with Cy3 dye as a reference and those of testes were labeled with Cy5 dye.
Project description:Microarray analysis of the gill tissues of WSSV infected shrimp (P. monodon) at different time intervals 6 hrs, 24 hrs, 48 hrs and moribund stage of post WSSV infection was carried out to identify differentially expressed genes in response to WSSV infection. The shrimps in WSSV challenege experiment were challenged through intra muscular route with known concentration of virus. The important immune genes identified would be further characterized by sequence analysis and gene expression profile would be validated by real time PCR One-color experiment,Organism: Penaeus monodon, Custom Penaeus monodon (Black Tiger Shrimp) 8x60k designed by Genotypic Technology Private Limited (AMADID: 041733), Labeling kit: Agilent Quick-Amp labeling Kit (p/n5190-0442)
Project description:Comparisons of gene expression profiles in hepatopancreas and ovary of domesticated broodstock at different ovarian maturation stages were made through a cDNA microarray in the black tiger shrimp (Penaeus monodon). Differentially expressed genes were identified through the microarray analysis, and the microarray results were confirmed by real-time PCR. Selected genes were further characterized.
Project description:Comparisons of gene expression profiles from testis of wild and domesticated male brooders were made through a cDNA microarray in the black tiger shrimp (Penaeus monodon). Differentially expressed genes were identified through the microarray analysis, and the microarray results were confirmed by real-time PCR. Selected genes were further characterized. A cDNA microarray was constructed from the EST libraries of P. monodon, consisting of 5,568 features. RNA samples were extracted from testes of pooled juveniles (4 month-old, pooled from n=114), broodstocks from West and East sides of Thailand, and domesticated broodstocks of 10-, 14-, and 18-month old. The RNA from testis of pooled juveniles was labeled with Cy3 dye as a reference and those of the broodstocks were labeled with Cy5 dye.
Project description:Comparisons of gene expression profiles between ovaries of before (day 0) and after eyestalk ablation (days 1, 4 and 7) of domesticated 14-month-old black tiger shrimp (Penaeus monodon) were made using a cDNA microarray. Differentially expressed genes were identified through the microarray analysis, and the microarray results were confirmed by real-time PCR. Selected genes were further characterized. A cDNA microarray consisting of 5,568 features was constructed from EST libraries of P. monodon. RNA samples were extracted from the ovaries of before (day 0) and after eyestalk ablation at days 1, 4 and 7 from shrimp from the Shrimp Genetic Improvement Center, Thailand. The RNA was converted into cDNA by indirect aminoallyl-cDNA labelling method (LabelStar Array, Qiagen). aa-cDNA from day 0 samples were coupled with Cy3 dye as a reference, and those from days 1, 4 and 7 were coupled with Cy5 dye.
Project description:Identification of testis-relevant genes using in silico analysis from testes ESTs and cDNA microarray in the black tiger shrimp (Penaeus monodon) A total of 4,803 ESTs from the testis library were unidirectionally sequenced and analyzed in silico using a customizable data analysis package, ESTplus. A total of 2,702 unique sequences from 424 contigs and 2,278 singletons were identified. Approximately, 1,134 sequences from the total of 2,702 unique sequences (contigs and singletons) are homologous to genes with known functions. Some of which (mago nashi, insulin-degrading enzyme, actin-depolymerizing factor) are related to testicular development in studies of other organisms. Moreover, the sequences were further characterized according to the gene ontology categories (41% biological process, 24% molecular function, 35% cellular component). By comparing with the EST libraries of other tissues, a total of 1,579 possible testis-specific ESTs were identified. Some of these testis-specific ESTs, such as Phospholipase A2 and Matrix Metalloproteinases, have functions relevant to testicular development. In addition, novel ESTs (621 ESTs) that have never been identified in any ESTs from the Penaeidae family were found in this study. Through cDNA microarray analysis, some of testis-specific ESTs were preferentially expressed in testes to ovary; one of which was a saposin homolog. Therefore, saposin was further characterized by real-time PCR and its full-length sequence was obtained by RACE-PCR. A cDNA microarray was constructed from the EST libraries of P. monodon, consisting of 5,376 features (1,125 features were amplified from testis EST libraries). RNA samples were extracted from testes and ovaries of juveniles (4 month-old, pooled from n=112 and n=98 for testes and ovaries, respectively) and broodstocks from Andaman Sea. The RNA from ovary was labeled with Cy3 dye as a reference and that from testes was labeled with Cy5 dye.
