scholarly journals Establishment of cDNA Library and EST Analysis from Leaves of Phyllanthus amarus

2014 ◽  
Vol 4 (1) ◽  
pp. 1-15 ◽  
Author(s):  
Sharmila Chattopadhyay
Keyword(s):  
Cdna Library ◽  
Phyllanthus Amarus ◽  
Est Analysis

Construction of cDNA library from Prunus campanulata leaves and preliminary expressed sequence tag (EST) analysis during cold stress

Biologia ◽  
2015 ◽  
Vol 70 (8) ◽  
Author(s):  
Ying-Hui Zhang ◽  
Jun-Dong Rong ◽  
Li-Guang Chen ◽  
Ling-Yan Chen ◽  
Tian-You He ◽  
...  
Keyword(s):  
Gene Expression ◽  
Low Temperature ◽  
Cold Stress ◽  
Cdna Library ◽  
Molecular Mechanisms ◽  
Expressed Sequence Tag ◽  
Go Annotation ◽  
Est Analysis ◽  
Young Leaves ◽  
Expressed Sequence

AbstractThe molecular mechanisms underlying cold-resistance in Prunus campanulata Maxim. (P. campanulata) are not fully understood. This study aimed to establish a full-length library and analyze expressed sequence tags (ESTs) to provide tools to investigate the mechanisms of P. campanulata growth at low temperatures. Based on the switching mechanism at 5’end of RNA transcript technology, a full-1ength cDNA library was generated from young leaves of P. campanulata after 72 h treatment at 1◦C, and a preliminary EST analysis was carried out. Quantitative reverse transcription polymerase chain reaction was used to assess the expression of selected cold-related genes. The cDNA library titer was 1.2 × 106 cfu/mL−1, with a recombinant rate of 96%. The average size of inserted cDNA fragments was 1.3 Kb. EST data revealed the existence of 834 clones representing a total of 667 unigenes, including 574 singletons and 93 contigs. Blast analysis identified 475 unigenes with known and putative functions. Based on similarity search and GO annotation, 84 unigenes were associated with “response to stimuli”, suggesting that cold stress induced significant alterations in gene expression in P. campanulata cultivated at 1◦C for 72 h. Interestingly, DRP, MYB, HSP, GPX and GA20-ox gene expression was significantly up-regulated in plants cultivated at low temperature, while transcript levels of TIL, CDPK were decreased. P. campanulata cultivating at low temperature express genes associated with “response to stimuli”, and in particular DRP, MYB, HSP, GPX and GA20-ox gene are up-regulated while TIL, CDPK are downregulated in response to low temperature-stress

scholarly journals Construction of cDNA Library and EST Analysis Related to Seed-hair Characteristics in Carrot

2013 ◽  
Vol 31 (6) ◽  
pp. 782-789 ◽  
Author(s):  
Gyu-Dong Oh ◽  
Eun-Jo Shim ◽  
Sang-Jin Jun ◽  
Young-Doo Park
Keyword(s):  
Cdna Library ◽  
Est Analysis ◽  
Seed Hair

Expressed sequence tag (EST) analysis of a Schistosomajaponicum cercariae cDNA library

Acta Tropica ◽  
2002 ◽  
Vol 82 (2) ◽  
pp. 215-224 ◽  
Author(s):  
Ming-Chiu Fung ◽  
Man-Tat Lau ◽  
Xiao-Guang Chen
Keyword(s):  
Cdna Library ◽  
Expressed Sequence Tag ◽  
Est Analysis ◽  
Expressed Sequence

EST analysis and annotation of transcripts derived from a trichome-specific cDNA library from Salvia fruticosa

2010 ◽  
Vol 29 (5) ◽  
pp. 523-534 ◽  
Author(s):  
Fani M. Chatzopoulou ◽  
Antonios M. Makris ◽  
Anagnostis Argiriou ◽  
Jörg Degenhardt ◽  
Angelos K. Kanellis
Keyword(s):  
Cdna Library ◽  
Est Analysis ◽  
Salvia Fruticosa

scholarly journals Total RNA Isolation and cDNA synthesis from Bixa orellana bark

2017 ◽  
Vol 7 ◽  
Author(s):  
Santosh P ◽  
Suresh B Arakera
Keyword(s):  
Gene Expression ◽  
Cdna Library ◽  
Rna Extraction ◽  
Rna Isolation ◽  
Natural Food ◽  
Bixa Orellana ◽  
Cdna Synthesis ◽  
Total Rna ◽  
Est Analysis ◽  
Study Gene Expression

<p><strong>Annatto is one of the important natural food colourant widely used in dairy industry. Quality RNA in large quantity is often required in the analysis of gene expression. RNA extraction from samples collected from woody plants is generally complex, and becomes the main limitation to study gene expression, particularly in crops like <em>Bixa orellana</em>. Standard RNA extraction protocols are time consuming, laborious and cannot be adapted for high throughput functional analysis. Therefore a simple and effective protocol for extraction of high quality total RNA from bark tissue of woody stem was achieved using the RNeasy plant mini kit (Qiagen, USA). The extracted RNA was successfully converted into double-stranded cDNA using the SMATer cDNA synthesis kit (Clontech, USA) which is based on the <span style="text-decoration: underline;">S</span>witching <span style="text-decoration: underline;">M</span>echanism <span style="text-decoration: underline;">A</span>t 5’ End of <span style="text-decoration: underline;">R</span>NA <span style="text-decoration: underline;">T</span>ranscript (SMART) technology. The integrity of the total RNA used for synthesizing double stranded cDNA was assessed agarose gel electrophoresis and by PCR. As expected, the PCR product contained the full coding sequence plus 69 and 196 bp of 5’ and 3’ UTRs respectively. The double-stranded cDNA was used successfully for creating a SSH cDNA library. The cDNA could also be useful for a number of other applications like cDNA library construction, EST analysis, RACE and Next Generation Sequencing (NGS).</strong></p>

An expression cDNA library for suppression cloning in yeast mutants, complementation of a yeast his4 mutant, and EST analysis from the symbiotic basidiomycete Hebeloma cylindrosporum

Genome ◽  
2003 ◽  
Vol 46 (2) ◽  
pp. 177-181 ◽  
Author(s):  
Daniel Wipf ◽  
Mariam Benjdia ◽  
Enno Rikirsch ◽  
Sabine Zimmermann ◽  
Mechthild Tegeder ◽  
...  
Keyword(s):  
Cdna Library ◽  
Gc Content ◽  
Expression Library ◽  
Significant Similarity ◽  
Hebeloma Cylindrosporum ◽  
Est Analysis ◽  
High Level ◽  
Fungal Genes ◽  
Yeast Mutants

An oriented expression library was constructed from the mycelia of the symbiotic model fungus Hebeloma cylindrosporum in the high-level yeast expression vector pDR196. DNA sequencing of ~500 expressed sequence tags (ESTs) showed that 15% correspond to known genes, two thirds contain sequences with unknown function, and the remaining 20% showed no significant similarity to any known genes. The ESTs had a GC content between 44 and 56%, with most of them having a GC content of 52–54%, which could be correlated with GC contents of fungal genes. The library was successfully used to identify the Hebeloma HIS4 gene by functional complementation of a yeast his4 mutant. Thus, the library may serve as a powerful tool for identification and characterization of mycorrhizal genes by EST analysis and for the identification of ectomycorrhizal genes by means of suppression cloning.Key words: ectomycorrhiza, functional cDNA library, Hebeloma cylindrosporum, yeast complementation.

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