Abstract:
OBJECTIVES:To investigate the effect of full-length fragment of DNA topoisomerase I gene (TOP1) matrix attachment regions (MARs) originating from the human genome on transgene expression in Chinese hamster ovary (CHO) cells and explore the underlying mechanisms. RESULTS:Results showed that TOP1 MAR cannot only enhance the transient and stable transgenic expression of enhanced green fluorescence protein (EGFP) but also increase long-term stability and ratio of positive colonies in transfected CHO cells with TOP1 MAR at the 5' or 3' ends of the EGFP expression cassette. Interestingly, the CHO cells were transfected with the 5',3' TOP1 MAR-containing vector featured the highest transient and stable expression, whereas those with the 3' TOP1 MAR-containing vector exhibited the most effective stability and ratio of positive colonies. We also observed that transgene copy numbers and mRNA of egfp gene were correlated with the expression levels of EGFP protein in polyclonal CHO cells. However, the heterogeneity of expression in monoclonal CHO cells was unaffected by transgene copy number. CONCLUSIONS:The findings may aid in the potential application of TOP1 MAR in expression enhancement of recombinant proteins in mammalian cells.
journal_name
Biotechnol Lettjournal_title
Biotechnology lettersauthors
Jia YL,Guo X,Wang XC,Wang TYdoi
10.1007/s10529-019-02673-7subject
Has Abstractpub_date
2019-07-01 00:00:00pages
701-709issue
6-7eissn
0141-5492issn
1573-6776pii
10.1007/s10529-019-02673-7journal_volume
41pub_type
杂志文章abstract::A novel two-step fermentation process was developed to enhance arachidonic acid (ARA) production by Mortierella alpina ME-1 in a 5 l fermentor. Agitation speed and aeration rate were adjusted from 180 to 40 rpm and from 0.6 to 1 vvm, respectively, after 5 days cultivation, to decrease physical damage to the mycelia an...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-008-9661-1
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-020-03002-z
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-010-0456-9
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abstract::Image analysis was used to measure the effect of Tween 40 on the morphology of Trichoderma harzianum. The percentage of pellets was maximal (93%) with Tween at 0.2 ml l(-1), whereas the maximal proportion of dispersed mycelia (40%) was with Tween at 1.6 ml l(-1). The particle median diameter decreased from 2 to 0.5 mm...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1023/b:bile.0000009458.58602.d9
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-007-9420-8
更新日期:2007-10-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-012-0996-2
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-006-9140-5
更新日期:2006-10-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1023/a:1024034508136
更新日期:2003-06-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-016-2101-8
更新日期:2016-08-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-010-0269-x
更新日期:2010-08-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-005-5528-x
更新日期:2006-02-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-005-3893-0
更新日期:2005-12-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 信件
doi:10.1007/s10529-014-1479-4
更新日期:2014-06-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-016-2099-y
更新日期:2016-07-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1023/b:bile.0000003990.27624.04
更新日期:2003-11-01 00:00:00
abstract::The transfer of the IncP-7 carbazole degradative plasmid pCAR1 from Pseudomonas putida SM1443 (derived from strain KT2440) into bacteria of river water samples was monitored using a reporter gene encoding red fluorescent protein (RFP). The number of transconjugants drastically increased in the presence of carbazole, a...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-007-9519-y
更新日期:2008-01-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-014-1631-1
更新日期:2014-12-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1023/B:BILE.0000045643.66758.9d
更新日期:2004-09-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章,评审
doi:10.1007/s10529-005-1521-7
更新日期:2005-03-01 00:00:00
abstract::N(omega)-2,2,4,6,7-Pentamethyldihydrobenzofuran-5-sulfonyl (N(omega)-Pbf)-protected oligoarginine was directly conjugated to poly(amino acid) derivatives modified with a long alkyl chain. The final concentration of conjugated peptides was easily controlled by the feed ratio of oligoarginine to polymer backbone and a f...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-005-7838-4
更新日期:2005-07-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-008-9730-5
更新日期:2008-09-01 00:00:00
abstract::Long-term cell recycle fermentations of Candida tropicalis were performed over 14 rounds of fermentation. The average xylitol concentrations, fermentation times, volumetric productivities and product yields for 14 rounds were 105 g l(-1), 333 h, 4.4 g l(-1) h(-1) and 78%, respectively, in complex medium; and 110 g l(-...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1023/b:bile.0000023019.02411.54
更新日期:2004-04-01 00:00:00
abstract:OBJECTIVE:The purpose of the article is to establish a quick enrichment and detection method using immunomagnetic beads and flow cytometry to analyze circulating tumor cells (CTCs) in the peripheral blood. RESULTS:After incubation with CD326-PE and CD45-APC antibodies, more than 60% MCF7 cells in M-Buffer could be det...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-020-03007-8
更新日期:2021-01-01 00:00:00
abstract::Virus-induced gene silencing is based on the sequence-specific degradation of RNA. Here, a gene silencing vector derived from EuMV-YP, named pEuMV-YP:ΔAV1, was used to silence ChlI and NPR1 genes in Nicotiana benthamiana. The silencing of the ChlI transcripts was efficient in the stems, petioles and leaves as reflecte...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-013-1146-1
更新日期:2013-05-01 00:00:00
abstract::The use of microRNAs (miRNAs) for improving the efficiency of recombinant protein production by CHO cells is gaining considerable interest for their ability to regulate entire molecular networks. Differential miRNA expression profiling and large-scale transient screening have been the prerequisite for the selection of...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-014-1751-7
更新日期:2015-04-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-017-2398-y
更新日期:2017-10-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-006-9215-3
更新日期:2007-01-01 00:00:00
abstract::Rhodococcus sp. CGMCC 4911 transformed 1,3-propanediol cyclic sulfate (1,3-PDS) and its derivatives into corresponding diols. Ethylene sulfate, glycol sulfide, 1,3-PDS, and 1,2-propanediol cyclic sulfate were effectively hydrolyzed with growing cells. (R)-1,2-Propanediol (>99 % e.e.) was obtained at 44 % yield with gr...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-014-1670-7
更新日期:2015-01-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章,评审
doi:10.1007/s10529-017-2303-8
更新日期:2017-06-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章,评审
doi:10.1007/s10529-015-1843-z
更新日期:2015-08-01 00:00:00