[Construction of cDNA library of protozoan ciliates Euplotes octocarinatus]

Wei Sheng Wu Xue Bao. 2010 Mar;50(3):316-21.
[Article in Chinese]

Abstract

The protein biosynthesis in cells is an open process cooperatively regulated by many protein factors and enzymes, which form a complicated protein network and signal transduction pathway for mRNA metabolism and protein translation.

Objective: To provide a platform for studying on the function of proteins involving in process of translation termination and the relationship among these proteins in ciliates.

Methods: We constructed the cDNA library of protozoan ciliates Euplotes octocarinatus strictly following the procedure of BD Matchmaker Library Construction & Screening kit from Clontech.

Results: We obtained a cDNA library of ciliates Euplotes, the titer of which was about 2.437 x 10(7) cfu/mL, which could suffice for functional gene screen. By using the class II polypeptide release factor (eRF3) as bait protein, we obtained some putative genes, including a partial cDNA putatively encoding RNA helicase.

Conclusion: This library will provide a convenient platform for identifying the functional genes in ciliates.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA, Protozoan / genetics
  • Euplotes / genetics*
  • Gene Library*
  • Peptide Termination Factors / genetics
  • Protozoan Proteins / genetics

Substances

  • DNA, Protozoan
  • Peptide Termination Factors
  • Protozoan Proteins
  • peptide-chain-release factor 3