Comparison of electrocompetencies of some commonly used laboratory strains of Escherichia coli

Comparison of electrocompetencies of some commonly used laboratory strains of Escherichia coli

During construction of genomic libraries from various bacteria, we experienced considerably different transformation effi ciencies among the strains of electrocompetent Escherichia coli cells and investigated the effect of type strain on effi ciency of transformation. Among the fi ve strains tested, Escherichia coli DH10B and TOP10 were found to be the most suitable strains for construction of genomic libraries in our laboratory. This study highlighted the fact that although these commercially available Escherichia coli strains are well characterized, they tend to differentiate and loose their transformation abilities. This may be due to the fact that the number of passages they have gone through affect their membrane structures and permeability. Therefore, every laboratory should test their strains for competency before they use them in their cloning studies.

___

  • [1] Glascock CB, Weickert MJ. 1998. Using chromosomal lacIQ1 to control expression of genes on high-copynumber plasmids in Escherichia coli. Gene. 223:221-31.
  • [2] Casali N. 2003. Escherichia coli host strains. Methods Mol Biol. 235:27-48.
  • [3] Mandel M, Higa, A. 1992. Calcium-dependent bacteriophage DNA infection. Biotechnology. 24:198-201.
  • [4] Cohen SN, Chang AC, Hsu L. 1972. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 8:2110-4.
  • [5] Hanahan D. 1983. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 166:557-80.
  • [6] Hanahan D, Jessee J, Bloom FR. 1991. Plasmid transformation of Escherichia coli and other bacteria. Methods Enzymol. 204:63-113.
  • [7] Inoue H, Nojima H, Okayama H. 1990. High effi ciency transformation of Escherichia coli with plasmids. Gene.96:23-8.
  • [8] Dower WJ, Miller JF, Ragsdale CW. 1988. High effi ciency transformation of Escherichia coli by high voltage electroporation. Nucleic Acids Res. 16:6127-45.
  • [9] Elvin S, Bingham AH. 1991. Electroporation-induced transformation of Escherichia coli: evaluation of a square waveform pulse. Lett Appl Microbiol. 12:39-42.
  • [10] Miller EM, Nickoloff JA. 1995. Escherichia coli electrotransformation. Methods Mol Biol. 47:105-13.
  • [11] Kasap M, Torol S, Gacar G, Budak F. 2006. Ethidium bromide spot test is a simple yet highly accurate method in determining DNA concentration. Turk J Med Sci; 36(6): 383-386.
  • [12] Tang X, Nakata Y, Li HO et al. 1994. The optimization of preparations of competent cells for transformation of E. coli. Nucleic Acids Res. 22:2857-8
Journal of Applied Biological Sciences-Cover
  • ISSN: 1307-1130
  • Yayın Aralığı: 6
  • Başlangıç: 2007
  • Yayıncı: Nobel Bilim ve Araştırma Merkezi
Sayıdaki Diğer Makaleler

Spatial analysis of yield and soil components in an onion field

Kh. M. DARWISH, A.A. Abd EL-KADER

Morphological, anatomical, palynological and cytological investigation on Silene urvillei Schott.(Caryophllaceae)

Ersin MİNARECİ, Kemal YILDIZ

Comparison of electrocompetencies of some commonly used laboratory strains of Escherichia coli

Murat KASAP, Gülçin GAÇAR, Sinem TOROL

Subcellular localization of Bm4, the protein product of ORF4 of Bombyx mori nucleopolyhedrovirus

Yan WU, Bing LI, Wu-Jie SU, Wei-De SHEN, Guang GAO, Wen-Bing WANG

Effects of high dose lead toxication on liver, kidneyse, heart, brain and blood in rabbits: An experimental study

Murat GÜZEL, Ahmet KOÇ, Ramazan DURGUT, Ramazan GÖNENCİ, Sefa ÇELİK, Ramazan BAL, M. Enes ALTUĞ, E. Özlem ATEŞOĞLU

Effect of applied iron on iron desorption in soil and uptake by wheat plants

F. A. HELLAL, A. K. AMER, A. M. ZAGHLOUL

Essential oil composition of Salvia aucheri Bentham var. canescens Bioiss. & Heldr. Described endemic species from Turkey

Ahmet DURAN, Ercüment Osman SARIHAN, Arif İPEK, Bilal GÜRBÜZ, Hayrettin KENDİR

Lignin degradation and acid precipitable polymeric lignin (APPL) accumulation by selected Streptomyces strains in submerged and solid state culture systems

A. Usame TAMER, Mustafa YAMAÇ

A Re-examination of the morphology of two endemic species of Centaurea L.(Asteraceae) from Turkey, with implications for taxonomy of the genus

Tuna UYSAL, Osman TUGAY

Production of poly-beta-hydroxybutyrate (PHB) in different media by Streptococcus thermophilus Ba21S Strain

Zehra Nur YÜKSEKDAĞ, Yavuz BEYATLI