Transformation of Anaplasma phagocytophilum.
Transformation of Anaplasma phagocytophilum.
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DOI：
10.1186/1472-6750-6-42
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发表时间：
2006-10-31
影响因子：
3.5
通讯作者：
Munderloh UG
中科院分区：
文献类型：
--
作者：
Felsheim RF;Herron MJ;Nelson CM;Burkhardt NY;Barbet AF;Kurtti TJ;Munderloh UG
壁虱传播的病原体会导致新出现的人畜共患病，包括挑剔的生物，如吞噬细胞性无浆体。由于它们专性的细胞内性质，诱变和转化的方法一直是不可用的。为了便于基因操作，我们用Himar1转座酶系统和临床无关的抗生素壮观霉素筛选，将吞噬细胞藻(Ap)转化为绿色荧光蛋白(GFP)。这些转化的细菌(GFP/AP)在人、猴子和扁虱细胞培养中以正常速度生长并发出明亮的荧光。对GFP/AP基因组DNA进行了分子鉴定，证实了转座和侧翼基因组插入位置。经腹腔注射GFP/AP的三只小鼠被感染，表现为外周血中性粒细胞中出现桑椹，并在培养中重新分离细菌。
Tick-borne pathogens cause emerging zoonoses, and include fastidious organisms such as Anaplasma phagocytophilum. Because of their obligate intracellular nature, methods for mutagenesis and transformation have not been available. To facilitate genetic manipulation, we transformed A. phagocytophilum (Ap) to express a green fluorescent protein (GFP) with the Himar1 transposase system and selection with the clinically irrelevant antibiotic spectinomycin. These transformed bacteria (GFP/Ap) grow at normal rates and are brightly fluorescent in human, monkey, and tick cell culture. Molecular characterization of the GFP/Ap genomic DNA confirmed transposition and the flanking genomic insertion locations were sequenced. Three mice inoculated with GFP/Ap by intraperitoneal injection became infected as demonstrated by the appearance of morulae in a peripheral blood neutrophil and re-isolation of the bacteria in culture.
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影响因子：
4.5
作者：
Dunning Hotopp JC;Lin M;Madupu R;Crabtree J;Angiuoli SV;Eisen JA;Seshadri R;Ren Q;Wu M;Utterback TR;Smith S;Lewis M;Khouri H;Zhang C;Niu H;Lin Q;Ohashi N;Zhi N;Nelson W;Brinkac LM;Dodson RJ;Rosovitz MJ;Sundaram J;Daugherty SC;Davidsen T;Durkin AS;Gwinn M;Haft DH;Selengut JD;Sullivan SA;Zafar N;Zhou L;Benahmed F;Forberger H;Halpin R;Mulligan S;Robinson J;White O;Rikihisa Y;Tettelin H
通讯作者：
Tettelin H
影响因子：
3.5
作者：
Felsheim RF;Herron MJ;Nelson CM;Burkhardt NY;Barbet AF;Kurtti TJ;Munderloh UG
通讯作者：
Munderloh UG
影响因子：
3.5
作者：
Barbet, AF;Agnes, JT;Palmer, GH
通讯作者：
Palmer, GH
影响因子：
6.5
作者：
ADES, EW;CANDAL, FJ;LAWLEY, TJ
通讯作者：
LAWLEY, TJ
影响因子：
2.1
作者：
Lukácová, M;Valková, D;Barák, I
通讯作者：
Barák, I