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Impact of heat shock step on bacterial transformation efficiency | ||
| Molecular Biology Research Communications | ||
| مقاله 6، دوره 5، شماره 4، اسفند 2016، صفحه 257-261 اصل مقاله (129.08 K) | ||
| نوع مقاله: Short communication | ||
| شناسه دیجیتال (DOI): 10.22099/mbrc.2016.3915 | ||
| نویسندگان | ||
| Maral Rahimzadeh1؛ Majid Sadeghizadeh* 2؛ Farhood Najafi3؛ Shahriar Arab4؛ Hamid Mobasheri5 | ||
| 1Department of Nanobiotechnology, School of Biological Sciences, Tarbiat Modares University, Tehran, Iran | ||
| 2Department of Molecular Genetics, School of Biological Sciences, Tarbiat Modares University, Tehran, Iran | ||
| 3Department of Resin and Additives, Institute for Color Science and Technology, Tehran, Iran | ||
| 4Department of Biophysics, School of Biological Sciences, Tarbiat Modares University, Tehran, Iran | ||
| 5Laboratory of Membrane Biophysics, Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran | ||
| چکیده | ||
| CaCl2 treatment followed by heat shock is the most common method for artificial transformation. Here, the cells were transformed using CaCl2 treatment either with heat shock (standard protocol) or without heat shock (lab protocol) to comprehend the difference in transformation efficiency. The BL21 strain of Escherichia coli (E. coli) was being susceptible using CaCl2 treatment. Some Cells were kept at -80 oC while the others were kept at 4 ̊C. Afterwards the susceptible cells were transformed using either standard or lab protocol. The transformation efficiency between cells experienced heat shock and those were not influenced by heat shock was almost the same. Moreover, regardless of transformation protocol, the cells kept at 4 ̊C were transformed more efficiently in compared to those were kept at -80 oC. | ||
| کلیدواژهها | ||
| E. coli؛ Artificial transformation؛ Heat shock؛ Transformation efficiency | ||
| مراجع | ||
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1. Chen I, Dubnau D. DNA uptake during bacterial transformation. Nat Rev Microbiol 2004;2:241-249. 2. Panja S, Saha S, Jana B, Basu T. Role of membrane potential on artificial transformation of E. coli with plasmid DNA. J Biotechnol 2006;127:14-20. 3. Divya Prakash G, Anish RV, Jagadeesh G, Chakravortty D. Bacterial transformation using micro-shock waves. Anal Biochem 2011;419:292-301. 4. Chung CT, Niemela SL, Miller RH. One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution. Proc Natl Acad Sci USA 1989;86:2172-2175. 5. Inoue H, Nojima H, Okayama H. High efficiency transformation of Escherichia coli with plasmids. Gene 1990;96:23-28. 6. van Die IM, Bergmans HE, Hoekstra WP. Transformation in Escherichia coli: studies on the role of the heat shock in induction of competence. J Gen Microbiol 1983;129:663-670. 7. Verkleij AJ, de Kruyff B, Ververgaert PH, Tocanne JF, van Deenen LL. The influence of pH, Ca2+ and protein on the thermotropic behaviour of the negatively charged phospholipid, phosphatidylglycerol. Biochim Biophys Acta 1974;339:432-437. 8. van Alphen L, Verkleij A, Burnell E, Lugtenberg B. 31P nuclear magnetic resonance and freeze-fracture electron microscopy studies on Escherichia coli. II. Lipopolysaccharide and lipopolysaccharide-phospholipid complexes. Biochim Biophys Acta 1980;597:502-517. 9. Gerritsen WJ, de Kruijff B, Verkleij AJ, de Gier J, van Deenen LL. Ca2+-induced isotropic motion and phosphatidylcholine flip-flop in phosphatidylcholine-cardiolipin bilayers. Biochim Biophys Acta 1980;598:554-560. 10. Mandersloot JG, Gerritsen WJ, Leunissen-Bijvelt J, van Echteld CJ, Noordam PC, de Gier J. Ca2+-induced changes in the barrier properties of cardiolipin/ phosphate-dylcholine bilayers. Biochim Biophys Acta 1981;640:106-113. 11. Burnell E, van Alphen L, Verkleij A, de Kruijff B. 31P nuclear magnetic resonance and freeze-fracture electron microscopy studies on Escherichia coli. I. Cytoplasmic membrane and total phospholipids. Biochim Biophys Acta 1980;597:492-501. | ||
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