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【答案】应助回帖
★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★
感谢参与，应助指数 +1
627455987: 金币+50, ★★★★★最佳答案, 非常感谢，近期较忙，忘记登录，抱歉 2017-08-23 20:57:40
心静_依然: LS-EPI+1, 感谢应助 2017-08-25 12:06:28
Expression of trehalose synthase in Pichia pastoris
Accession number: 20140417231137
Authors: Wang, Rui-Ming 1 ; Li, Zhong-Kui 1 ; Wang, Teng-Fei 1 ; Liu, Hong-Juan 1 ; Li, Pi-Wu 1
Author affiliation : 1 Department of food and Biology Engineering, QILU university of technology, Jinan 250353, China
Corresponding author: Wang, R.-M.
Source title: Modern Food Science and Technology
Abbreviated source title: Mod. Food Sci. Technol.
Volume: 29
Issue: 11
Issue date: 2013
Publication Year: 2013
Pages: 2575-2579
Language: Chinese
ISSN: 16739078
Document type: Journal article (JA)
Publisher: South China University of Technology, Guangzhou, 510640, China
Abstract: The expression vector was constructed from trehalose synthase (TreS) gene(AE015451.1) of Pseudomonas putida to express in Pichia pastoris. TreS gene was amplified by PCR and double digested by EcoRI/XbaI, then linked to the pPICZaA which digested with same enzymes. The gene was identified, sequenced and transformed into pichia pastoris GS115 by electroporation. Positive colonies harboring target genes were screened out by the YPD medium with Zeocin. The genome of positive colonies were extracted and a same size band with target gene was abtained by using PCR which illustrated target gene had successfully transferred to Pichia pastoris. SDS-PAGE and HPLC results showed that the recombinant enzyme had a molecular of 76 ku band which was consistent with the predicted molecular mass, and it had the ability to catalyze the conversion of maltose into trehalose. The recombinant pPICZaA-TreS plasmid successfully constructed and integrated into Pichia pastoris genome and expressed as expected.
Number of references: 14
Main heading: Gene expression
Controlled terms: Yeast
Uncontrolled terms: Electroporation - Expression vectors - Pichia Pastoris - Pichia pastoris gs115 - Pseudomonas putida - Recombinant enzymes - Trehalose - Trehalose synthase
Classification code: 461.8.1Genetic Engineering - 801.2Biochemistry
Database: Compendex
Compilation and indexing terms, © 2017 Elsevier Inc. |
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