Quantified High-Throughput Screening of Escherichia coli Producing Poly(3-hydroxybutyrate) Based on FACS

被引：29

作者：

Lee, Jae Hyung ^{[1
]}

Lee, Seung Hwan ^{[2
]}

Yim, Sung Sun ^{[1
]}

Kang, Kyoung-Hee ^{[2
,5
,6
]}

Lee, Sang Yup ^{[3
,4
]}

Park, Si Jae ^{[5
,6
]}

Jeong, Ki Jun ^{[1
]}

机构：

[1] Korea Adv Inst Sci & Technol, Dept Chem & Biomol Engn, Program BK21, Taejon 305701, South Korea

[2] Korea Res Inst Chem Technol, Div Convergence Chem, Res Ctr Biobased Chem, Ind Biochem Res Grp, Taejon 305600, South Korea

[3] Korea Adv Inst Sci & Technol, Ctr Syst & Synthet Biotechnol, Dept Chem & Biomol Engn, Metab & Biomol Engn Natl Res Lab,Program BK21, Taejon 305701, South Korea

[4] Korea Adv Inst Sci & Technol, Inst BioCentury, Taejon 305701, South Korea

[5] Myongji Univ, Dept Environm Engn & Energy, Yongin 449728, Gyeonggido, South Korea

[6] Myongji Univ, Dept Energy & Biotechnol, Yongin 449728, Gyeonggido, South Korea

来源：

APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY | 2013年 / 170卷 / 07期

关键词：

FACS; BODIPY; Nile red; Poly(3-hydroxybutyrate); Escherichia coli; BETA-HYDROXYBUTYRIC ACID; PHA SYNTHASE; NILE RED; FLOW-CYTOMETRY; POLY-3-HYDROXYBUTYRATE; POLYHYDROXYALKANOATES; EVOLUTION; PROTEOME;

D O I：

10.1007/s12010-013-0311-2

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Here, we report on a highly sensitive method for the detection of P(3HB) accumulation in Escherichia coli cells based on the automated flow cytometry system using fluorescent dyes. E. coli containing P(3HB) were stained with either BODIPY or Nile red fluorescent dye, and their staining properties were analyzed under a variety of conditions. Compared with Nile red, BODIPY was much more sensitive in staining P(3HB) and overall demonstrated a more rapid staining of cells, a greater resistance to photobleaching, and greater cell viability. In addition, we also successfully monitored heterogeneity in P(3HB) accumulation within a cell population using BODIPY staining and flow cytometry. We believe this optimized staining method using BODIPY in combination with screening by high-speed flow cytometer will be helpful in the engineering of host cells toward an enhanced production of bioplastics.

引用

页码：1767 / 1779

页数：13

共 29 条

[21] New insights into the solubilization of Bodipy dyes [J].