Stage-specific expressed sequence tags obtained during preimplantation bovine development by differential display RT-PCR and suppression subtractive hybridization

2002 ◽  
Vol 22 (12) ◽  
pp. 1135-1142 ◽  
Author(s):  
Siriluck Ponsuksili ◽  
David Tesfaye ◽  
Nermin El-Halawany ◽  
Karl Schellander ◽  
Klaus Wimmers
Keyword(s):  
Suppression Subtractive Hybridization ◽  
Differential Display ◽  
Expressed Sequence Tags ◽  
Subtractive Hybridization ◽  
Rt Pcr ◽  
Expressed Sequence

scholarly journals Isolation of biosynthesis related transcripts of 2,3,5,4'-tetrahydroxy stilbene-2-O-β-D-glucoside from Fallopia multiflora by suppression subtractive hybridization

2014 ◽  
Vol 83 (2) ◽  
pp. 147-157 ◽  
Author(s):  
Wei Zhao ◽  
Shujing Sheng ◽  
Zhongyu Liu ◽  
Di Lu ◽  
Kuanpeng Zhu ◽  
...  
Keyword(s):  
Suppression Subtractive Hybridization ◽  
Expressed Sequence Tags ◽  
Digital Gene Expression ◽  
Subtractive Hybridization ◽  
Full Length ◽  
Specific Difference ◽  
Protein Databases ◽  
Organ Specific ◽  
Rapid Amplification ◽  
Expressed Sequence

2,3,5,4'-tetrahydroxy stilbene-2-O-ß-D-glucoside (THSG) exerts multiple pharmacodynamic actions, found in <em>Fallopia multiflora</em>, but the biosynthesis pathway of THSG is still unclear. To clear this ambiguity, we constructed suppression subtractive hybridization (SSH) libraries to screen the genes involved in THSG biosynthesis from two <em>F. multiflora</em> varieties, which vary significantly in THSG content. Twelve non-redundant differentially expressed sequence tags were obtained and the full lengths of 4 unreported fragments were amplified by rapid amplification of cDNA ends. We totally got 7 full-length transcripts, and all of them were aligned to the transcriptome and digital gene expression tag profiling database of four <em>F. multiflora</em> tissues (root, stem and leaf from Deqing <em>F. multiflora</em> and another root from Chongqing <em>F. multiflora</em>; data unpublished) using local BLAST. The results showed that there was a significant, organ specific difference in the expression of fragments and full-length sequences. All the sequences were annotated by aligning to nucleotide and protein databases. Kyoto Encyclopedia of Genes and Genomes pathway analysis indicated that THSG biosynthesis was correlated with multiple life activities.

Isolation, identification and expression profile of fourteen differentially expressed sequence tags in the longissimus dorsi muscle tissues from Meishan, Meishan×Large White cross and Large White pigs

2007 ◽  
Vol 4 (1) ◽  
pp. 9-14
Author(s):  
Liu Yong-Gang ◽  
Xiong Yuan-Zhu ◽  
Zuo Bo ◽  
Jiang Si-Wen ◽  
Deng Chang-Yan ◽  
...  
Keyword(s):  
Expression Profile ◽  
Differential Display ◽  
Expressed Sequence Tags ◽  
Differentially Expressed ◽  
Longissimus Dorsi ◽  
Large White ◽  
Longissimus Dorsi Muscle ◽  
Dorsi Muscle ◽  
Muscle Tissues ◽  
Expressed Sequence

AbstractIn order to detect the molecular mechanism of heterosis in pigs, an mRNA differential display (DD) technique was performed to investigate the differences in gene expression in the longissimus dorsi muscle tissues from Meishan, Meishan×Large White cross and Large White pigs. Fourteen expressed sequence tags (ESTs), differentially expressed between the hybrid and purebred pigs, were isolated and identified through semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). Nucleotide sequence analysis revealed that the 14 ESTs are not homologous to any of the known genes or ESTs. These novel ESTs were then deposited in the GenBank database. Tissue expression profile analysis showed that the ESTs were expressed in most tissues, including heart, spleen, liver, kidney, small intestine, ovary and lung, and this also implied that these genes must be important for the life process. Our results indicate the diversity of differential display of genes between the hybrids and purebreds in the Meishan×Large White cross combination. Results also suggest that heterosis in pigs might be derived from the differential expression of many indispensable genes in specific life phases.

scholarly journals IDENTIFICACIÓN DE cDNAs DE Agave tequilana Weber Var. azul SIMILARES A FUROSTANOL GLICÓSIDO 26-O-β-GLUCOSIDASAS

2019 ◽  
Vol 42 (4) ◽  
pp. 439-447
Author(s):  
Zurisadaí Monroy-González ◽  
Aída Martínez-Hernández
Keyword(s):  
Expressed Sequence Tags ◽  
Asparagus Officinalis ◽  
Agave Tequilana ◽  
Rt Pcr ◽  
Expresión Génica ◽  
Expressed Sequence

La furostanol glicósido 26-O-β-glucosidasa (F26G) participa en el último paso de la biosíntesis de saponinas esteroidales, lo que escinde la glucosa unida al hidroxilo del C26 del furostanol glucósido para permitir la formación del espirostano. A pesar de la existencia de numerosos estudios que muestran la diversidad estructural y actividad biológica de las saponinas de plantas, así como su reconocido potencial biotecnológico y farmacológico, pocas enzimas participantes en la ruta de biosíntesis de las saponinas esteroidales han sido estudiadas y sólo una enzima nativa tipo F26G ha sido purificada y caracterizada funcionalmente. En este estudio, mediante búsqueda por BLASTX en una base de datos de Agave tequilana Weber var. azul (Agave DB), se identificaron 46 ESTs (Expressed Sequence Tags) de DNA complementario (cDNAs) similares a la F26G de Asparagus officinalis, la mayoría provienen de cDNAs clonados a partir de tejidos florales (anteras y ovarios) o raíz; ambos órganos son productores de saponinas. Entre ellos, se identificaron seis ESTs cuyo alineamiento indica que representan a tres cDNAs diferentes entre sí. Estos ESTs se registraron en la base de datos dbEST de NCBI (National Center for Biotechnology Information), el cual fue el primer registro de secuencias nativas de cDNAs potencialmente codificantes similares a F26G en Agave. El tamaño de los cDNAs clonados y su alineamiento hacia el lado 5’ de las F26G conocidas sugieren que son cDNAs completos. Se diseñaron iniciadores diferenciales específicos para cada tipo de cDNA y se analizó por RT-PCR su expresión en tejidos vegetativos de A. tequilana. Se establecieron protocolos de PCR cuantitativo (qPCR) para estudios posteriores de regulación de su expresión génica. La información aquí reportada servirá como base para estudiar la función, regulación y actividad enzimática de las enzimas clonadas de Agave similares a F26G.

scholarly journals Generation and Analysis of Expressed Sequence Tags fromChimonanthus praecox(Wintersweet) Flowers for Discovering Stress-Responsive and Floral Development-Related Genes

2012 ◽  
Vol 2012 ◽  
pp. 1-13 ◽  
Author(s):  
Shunzhao Sui ◽  
Jianghui Luo ◽  
Jing Ma ◽  
Qinlong Zhu ◽  
Xinghua Lei ◽  
...  
Keyword(s):  
Gene Ontology ◽  
Real Time ◽  
Expressed Sequence Tags ◽  
Floral Development ◽  
Expression Patterns ◽  
Read Length ◽  
Rt Pcr ◽  
Protein Database ◽  
Blast Analysis ◽  
Expressed Sequence

A complementary DNA library was constructed from the flowers ofChimonanthus praecox, an ornamental perennial shrub blossoming in winter in China. Eight hundred sixty-seven high-quality expressed sequence tag sequences with an average read length of 673.8 bp were acquired. A nonredundant set of 479 unigenes, including 94 contigs and 385 singletons, was identified after the expressed sequence tags were clustered and assembled. BLAST analysis against the nonredundant protein database and nonredundant nucleotide database revealed that 405 unigenes shared significant homology with known genes. The homologous unigenes were categorized according to Gene Ontology hierarchies (biological, cellular, and molecular). By BLAST analysis and Gene Ontology annotation, 95 unigenes involved in stress and defense and 19 unigenes related to floral development were identified based on existing knowledge. Twelve genes, of which 9 were annotated as “cold response,” were examined by real-time RT-PCR to understand the changes in expression patterns under cold stress and to validate the findings. Fourteen genes, including 11 genes related to floral development, were also detected by real-time RT-PCR to validate the expression patterns in the blooming process and in different tissues. This study provides a useful basis for the genomic analysis ofC. praecox.

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