
Tn-seq轉(zhuǎn)座子測(cè)序技術(shù)服務(wù)介紹:
對(duì)基因功能的了解落后于基因發(fā)現(xiàn)阻礙了我們對(duì)微生物表型遺傳基礎(chǔ)的理解。 大規(guī)模并行測(cè)序與傳統(tǒng)的轉(zhuǎn)座子誘變相結(jié)合的轉(zhuǎn)座子測(cè)序(Tn-seq),可以以高通量的方式鑒定推定的基因功能。
轉(zhuǎn)座子測(cè)序需要構(gòu)建一個(gè)轉(zhuǎn)座子插入文庫,其中大多數(shù)或所有非必需基因(non-essential genes)都包含插入物,然后在特定的體外或體內(nèi)(宿主感染)條件下培養(yǎng)。通過測(cè)序確定實(shí)驗(yàn)的開始和結(jié)束時(shí)種群中每個(gè)突變體的相對(duì)頻率。從這些數(shù)據(jù)可以量化每個(gè)基因在每種情況下的適應(yīng)性貢獻(xiàn),分析出哪些基因是必需基因(essential genes)及推測(cè)基因與特定表型的相關(guān)性。例如,通過比較突變豐度來鑒定抗生素抗性基因。

沃森服務(wù)流程
沃森生物提供的轉(zhuǎn)座子測(cè)序服務(wù)接收構(gòu)建好的突變體庫菌體樣本,或提取好的突變體庫菌體基因組DNA,進(jìn)行NGS建庫測(cè)序及后續(xù)的分析工作。

數(shù)據(jù)分析流程

可視化結(jié)果

參考文獻(xiàn)
[1] Tim van Opijnen, et al. Tn-seq: high-throughput parallel sequencing for fitness and genetic interaction studies in microorganisms. Nature methods, 6, 767-772 (2009).
[2] Tim van Opijnen, Andrew Camilli. Transposon insertion sequencing: a new tool for systems-level analysis of microorganisms. Nature Reviews, 11, 435-442 (2013).
[3] Michael C. Chao, et al. The design and analysis of transposon insertion sequencing experiments. Nature Reviews, 14, 119-128 (2016).
[4] Larry A. Gallagher. Methods for Tn-Seq Analysis in Acinetobacter baumannii. Methods in Molecular Biology, 1946, 115-134 (2019).
[5] Andrew K. Fenton, et al. CozE is a member of the MreCD complex that directs cell elongation in Streptococcus pneumonia. Nature Microbiology, 2, 16237 (2016).
[6] Almudena Gonzalez-Mula, et al. The biotroph Agrobacterium tumefaciens thrives in tumors by exploiting a wide spectrum of plant host metabolites. New Phytologist, 222: 455–467(2019).