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天蚕抗菌肽XJ（一种来自鳞翅目蚕蛾科昆虫的生物活性抗菌肽）在（此处原文缺失相关宿主信息）中的表达与特性分析

Expression and characterization of cecropinXJ, a bioactive antimicrobial peptide from (Bombycidae, Lepidoptera) in .

作者信息

Xia Lijie, Zhang Fuchun, Liu Zhongyuan, Ma Ji, Yang Jianhua

机构信息

Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi, Xinjiang 830046, P.R. China ;

出版信息

Exp Ther Med. 2013 Jun;5(6):1745-1751. doi: 10.3892/etm.2013.1056. Epub 2013 Apr 9.

DOI:10.3892/etm.2013.1056
PMID:23837066
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3702707/
Abstract

Insect antimicrobial peptides (AMPs) have a broad antimicrobial spectrum. To aid the characterization of the gene function and further applications, we cloned the gene of cecropinXJ into the prokaryotic expression vector pET32a and expressed cecropinXJ in BL2l (DE3). Following induction by isopropyl-β-D-thiogalactoside (IPTG), a 25 kDa fusion peptide of cecropinXJ with a tagged thioredoxin (Trx) protein was highly expressed in . The yield was 10 mg/l culture medium following purification on nickel-nitrilotriacetic acid (Ni-NTA) metal affinity chromatography matrices. The purified recombinant antibacterial peptide, cecropinXJ, retained a high stability against over a temperature range from 4 to 100°C and a pH range from pH 2.0 to 12.0. The minimum inhibitory concentration (MIC) of the fusion protein against was 0.4 M. The recombinant cecropinXJ is also cytotoxic to several types of human cancer cells.

摘要

昆虫抗菌肽（AMPs）具有广泛的抗菌谱。为了有助于基因功能的表征和进一步应用，我们将天蚕素XJ基因克隆到原核表达载体pET32a中，并在BL2l（DE3）中表达天蚕素XJ。用异丙基-β-D-硫代半乳糖苷（IPTG）诱导后，天蚕素XJ与带标签的硫氧还蛋白（Trx）蛋白的25 kDa融合肽在[具体表达宿主]中高表达。在镍-次氮基三乙酸（Ni-NTA）金属亲和色谱基质上纯化后，产量为10 mg/升培养基。纯化后的重组抗菌肽天蚕素XJ在4至100°C的温度范围和pH 2.0至12.0的pH范围内对[具体物质]保持高度稳定性。融合蛋白对[具体物质]的最小抑菌浓度（MIC）为0.4 M。重组天蚕素XJ对几种类型的人类癌细胞也具有细胞毒性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c24/3702707/1ad03a0aa274/ETM-05-06-1745-g05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c24/3702707/0e669ebc9136/ETM-05-06-1745-g00.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c24/3702707/454c60f5ac52/ETM-05-06-1745-g01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c24/3702707/d47a3bb94d65/ETM-05-06-1745-g02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c24/3702707/23fb14f23ac8/ETM-05-06-1745-g03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c24/3702707/830c3b2681f6/ETM-05-06-1745-g04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c24/3702707/1ad03a0aa274/ETM-05-06-1745-g05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c24/3702707/0e669ebc9136/ETM-05-06-1745-g00.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c24/3702707/454c60f5ac52/ETM-05-06-1745-g01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c24/3702707/d47a3bb94d65/ETM-05-06-1745-g02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c24/3702707/23fb14f23ac8/ETM-05-06-1745-g03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c24/3702707/830c3b2681f6/ETM-05-06-1745-g04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c24/3702707/1ad03a0aa274/ETM-05-06-1745-g05.jpg

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