羊莫氏巴贝斯虫细菌人工染色体（BAC）文库的构建

doi:10.11843/j.issn.0366-6964.2014.08.017

畜牧兽医学报 ›› 2014, Vol. 45 ›› Issue (8): 1324-1329.doi: 10.11843/j.issn.0366-6964.2014.08.017

羊莫氏巴贝斯虫细菌人工染色体（BAC）文库的构建

张浩浩^1,3，陈龙欣^2,3，曹阳³，陈瑛³，关贵全^1*，马润林^2,3*

（1.中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室甘肃省动物寄生虫病重点实验室,兰州 730046；2.郑州师范学院，郑州 450044；3.中国科学院遗传与发育生物学研究所分子发育生物学国家重点实验室，北京 100101）

收稿日期:2014-01-25 出版日期:2014-08-23 发布日期:2014-08-23
通讯作者: 关贵全，副研究员，E-mail：guanguiquan@163.com；马润林，研究员，E-mail：rlma@genetics.ac.cn
作者简介:张浩浩（1988-），男，河南济源人，硕士生，主要从事寄生虫分子生物学研究，E-mail：haohaozhang2012@163.com，Tel：0931-8342681
基金资助:
家畜疫病病原生物学国家重点实验室开放课题（SKLVEB2012KFKT001）

Construction of Bacterial Artificial Chromosome (BAC) Library of Ovine Babesia motasi

ZHANG Hao-hao^1,3，CHEN Long-xin^2,3，CAO Yang³，CHEN Ying³，GUAN Gui-quan^1*，MA Run-lin^2,3*

（1.State Key Laboratory of Veterinary Etiological Biology/Key Laboratory of Veterinary Parasitology of Gansu Province/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences，Lanzhou 730046，China；2.Zhengzhou Normal University,Zhengzhou 450044，China；3.Institute of Genetics and Developmental Biology，Chinese Academy of Sciences，Beijing 100101，China)

Received:2014-01-25 Online:2014-08-23 Published:2014-08-23

摘要/Abstract

摘要：

为丰富巴贝斯虫基因组信息，选用感染中国羊的莫氏巴贝斯虫临潭株(Babesia motasi Lintan)为对象，将纯化的红细胞阶段虫体裂殖子包埋于低熔点琼脂糖，用蛋白酶K进行消化处理，得到巴贝斯虫基因组DNA，利用脉冲场凝胶电泳（PFGE)分离大片段基因组DNA，将其与CopyControl pCC1BAC Cloning-ready Vector进行连接并转化到EPI300^TM Escherichia coli 感受态细胞，成功构建了含有34 560 个克隆的莫氏巴贝斯虫临潭株基因组细菌人工染色体（BAC）文库；文库的平均插入片段大小为70 kb，基因组覆盖度为×137，空载率均低于3%，且克隆稳定。该BAC文库的构建，为进一步绘制精细物理图谱、基因组测序、进化及利用比较基因组学筛选致病基因等方面的研究奠定了基础。

Abstract:

To enrich the genomic information，the merozoites of ovine Babesia motasi Lintan were embedded in low-melting-point (LMP) agarose and subsequently digested with proteinase K to get their high-quality megabase size genomic DNA.Then the large size DNA fragments isolated by pulse-field gel electrophoresis (PFGE) were ligated to CopyControl pCC1BAC Cloning-ready Vector.The present results showed that the bacterial artificial chromosome (BAC) library containing 34 560 clones was successfully constructed by transformation of the ligated DNA into EPI300^TM Escherichia coli.In B.motasi Lintan BAC library，the average length of the insert DNA fragment was 70 kb and covers ×137 B.motasi Lintan genome.The BAC library in which the empty clone was less than 3% and the clones were stable should contribute to fine physical mapping，genome sequencing，and evolution and virulence genes derived from comparative genomics of Babesia species and so on.

中图分类号:

S852.723

张浩浩，陈龙欣，曹阳，陈瑛，关贵全，马润林. 羊莫氏巴贝斯虫细菌人工染色体（BAC）文库的构建[J]. 畜牧兽医学报, 2014, 45(8): 1324-1329.

ZHANG Hao-hao，CHEN Long-xin，CAO Yang，CHEN Ying，GUAN Gui-quan，MA Run-lin. Construction of Bacterial Artificial Chromosome (BAC) Library of Ovine Babesia motasi[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(8): 1324-1329.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/10.11843/j.issn.0366-6964.2014.08.017

https://www.xmsyxb.com/CN/Y2014/V45/I8/1324

参考文献

［1］YIN H,LU W,LUO J X.Babesiosis in China ［J］.Trop Anim Health Prod,1997，29(4):11-15.
［2］LIU A H，YIN H，GUAN G Q，et al.At least two genetically distinct large Babesia species infective to sheep and goats in China［J］.Vet Parasitol,2007,147(3-4):246-251.
［3］GUAN G Q,MA M L，MOREAU E，et al.A new ovine Babesia species transmitted by Hyalomma anatolicum anatolicum［J］.Exp Parasitol,2009，122(4):261-267.
［4］NIU Q L，LUO J X，GUAN G Q，et al.Differentiation of two ovine Babesia based on the ribosomal DNA internal transcribed spacer (ITS) sequences［J］.Exp Parasitol,2009，121(1):64-68.
［5］GUAN G Q,YIN H,LUO J X，et al.Transmission of Babesia sp to sheep with field-collected Haemaphysalis qinghaiensis［J］.Parasitol Res,2002，88(13 Suppl 1):S22- S24.
［6］GUAN G Q,CHAUVIN A,YIN H,et al.BQ1 (Lintan) and B.divergens in sheep depends on the production of IFNgamma and IL10［J］.Parasite Immunol，2010,32(2):143-152.
［7］GUAN G Q,MA M L,LIU A H，et al.Continuous IN VITRO cultivation of a recently identified BABESIA that infects small ruminants in China［J］.Vet Parasitol，2012,187(3-4):371-378.
［8］FIGUEROA J V,BUENING G M,KINDEN D A.Purification of the erythrocytic stages of BABESIA BIGEMINA from cultures［J］.Parasitol Res,1990,76(8):675-680.
［9］BHUSHAN C,MÜLLER I,FRIEDHOFF K T.Enrichment of Babesia caballi-infected erythrocytes from microaerophilous stationary-phase cultures using Percoll gradients ［J］.Parasitol Res,1991,77(2):177-179.
［10］BIRREN B，GREEN E D，KLAPHOLZ S，et al.Analyzing DNA：A laboratory manual［M］.NewYork：Cold Spring Harbor Laboratory Press，1997.
［11］ZHANG H B,SCHEURING C F,ZHANG M，et al.Construction of BIBAC and BAC libraries from a variety of organisms for advanced genomics research［J］.Nat Protoc,2012,7(3):479-499.
［12］TAO Q,ZHANG H B.Cloning and stable maintenance of DNA fragments over 300 kb in Escherichia coli with conventional plasmid-based vectors［J］.Nucleic Acids Res,1998，26(21):4901-4909.
［13］GARDNER M J,BISHOP R,SHAH T，et al.Genome sequence of Theileria parva，a bovine pathogen that transforms lymphocytes［J］.Science,2005,309(5731):134-137.
［14］PAIN A,RENAULD H，BERRIMAN M,et al.Genome of the host-cell transforming parasite Theileria annulata compared with T.parva［J］.Science,2005,309(5731):131-133.
［15］BRAYTON K A,LAU A O,HERNDON D R，et al.Genome sequence of Babesia bovis and comparative analysis of apicomplexan hemoprotozoa［J］.PLoS Pathog,2007,3(10):1401-1413.
［16］CORNILLOT E,HADJ-KADDOUR K,DASSOULI A，et al.Sequencing of the smallest Apicomplexan genome from the human pathogen Babesia microti［J］.Nucleic Acids Res,2012,40(18):9102-9114.
［17］贺兰.东方巴贝斯虫新型诊断方法的建立和基因组序列测定与分析［D］.武汉：华中农业大学，2011.

[1]	肖洁, 罗跃军, 陈浩, 蒲家艳, 何维, 熊常明, 郝哥, 任永军, 杨光友. 兔斯氏艾美耳球虫、肠艾美耳球虫和大型艾美耳球虫PCR检测靶标的筛选与单一、多重直接PCR检测方法的建立[J]. 畜牧兽医学报, 2023, 54(10): 4327-4337.
[2]	郑若愚, 肖洁, 白鑫, 陈浩, 蒲家艳, 任永军, 杨光友. 大型艾美耳球虫重组蛋白GAPDH对兔的免疫保护效果评价[J]. 畜牧兽医学报, 2023, 54(10): 4338-4349.
[3]	郑若愚, 任永军, 肖洁, 白鑫, 蒲家艳, 陈浩, 杨光友. 斯氏艾美耳球虫3-磷酸甘油醛脱氢酶重组蛋白对兔的免疫保护效果评价[J]. 畜牧兽医学报, 2023, 54(6): 2581-2595.
[4]	王礼跃, 冯茜茜, 蔡为民, 刘丹丹, 候照峰, 康喜龙, 张知之, 范雪莲, 朱玉, 许金俊, 潘志明, 陶建平. 鸡毒害艾美耳球虫ApiAP2转录因子的克隆、表达与虫体内定位[J]. 畜牧兽医学报, 2022, 53(12): 4379-4388.
[5]	赵保才, 赵帅阳, 关贵全, 罗建勋, 曹天行, 张志刚, 石苗, 刘军龙, 赵洪喜. 基于转录组数据分析环形泰勒虫对诱导宿主细胞转化相关基因的影响[J]. 畜牧兽医学报, 2022, 53(12): 4398-4409.
[6]	尹艳玲, 黄爽, 姚倩, 吴江平, 郭浩晨, 杨新, 宋军科, 赵光辉. 环状RNA ciRS-7靶向miR-219a-5p影响微小隐孢子虫体外增殖的机制[J]. 畜牧兽医学报, 2022, 53(11): 3989-3999.
[7]	肖克, 陈婷, 赵其平, 朱顺海, 董辉, 刘曼玉, 于钰, 黄兵, 韩红玉. 柔嫩艾美耳球虫含HD结构域蛋白特性和功能初步研究[J]. 畜牧兽医学报, 2022, 53(8): 2608-2620.
[8]	叶状, 王乐乐, 汪飞燕, 刘悦, 彭月梅, 宿世杰, 候照峰, 许金俊, 陶建平, 刘丹丹. 毒害艾美耳球虫氧化还原酶EnOXIO1的原核表达与定位分析[J]. 畜牧兽医学报, 2022, 53(5): 1553-1561.
[9]	焦宇洲, 杨欢欢, 卢垚, 史聪聪, 徐晓娟, 蔡旭旺. 屎肠球菌对感染柔嫩艾美耳球虫肉鸡生长性能和肠道屏障的影响[J]. 畜牧兽医学报, 2022, 53(4): 1210-1219.
[10]	罗跃军, 任永军, 白鑫, 陈浩, 蒲家艳, 郑若愚, 肖洁, 杨光友. 斯氏艾美耳球虫重组表面抗原SAG13和SAG14对兔的免疫保护效果初步观察[J]. 畜牧兽医学报, 2022, 53(3): 883-893.
[11]	梁珊珊, 朱顺海, 赵其平, 黄兵, 董辉, 于钰, 王青杰, 余水兰, 王海霞, 韩红玉. 柔嫩艾美耳球虫延伸因子1α特性与功能初步分析[J]. 畜牧兽医学报, 2020, 51(12): 3111-3121.
[12]	宋瑞其, 呼尔查, 翟雪洁, 樊新丽, 李敏, 张梦圆, 宋晶晶, 阿尔曼·海热, 吾力江·卡马力, 巴音查汗·盖力克. 马泰勒虫和驽巴贝斯虫半胱氨酸蛋白酶截短基因的克隆表达及其生物信息学分析[J]. 畜牧兽医学报, 2020, 51(10): 2547-2556.
[13]	杨守深, 孙晓双, 邱云飞, 杨飞, 唐杰, 连文健, 黄翠琴, 邱泓铨. 猪囊等孢球虫孢子化与未孢子化卵囊的差异表达蛋白质组学分析[J]. 畜牧兽医学报, 2020, 51(9): 2216-2226.
[14]	王璐, 朱顺海, 赵其平, 黄兵, 韩红玉, 刘桂玲, 李志行, 赵焕之, 董辉. 宿主GAPDH对柔嫩艾美耳球虫子孢子入侵细胞的影响[J]. 畜牧兽医学报, 2019, 50(10): 2088-2096.
[15]	王盼举, 樊新丽, 张梦圆, 宋晶晶, 李敏, 吾力江, 巴音查汗. 地方株马驽巴贝斯虫Bc48截短体单克隆抗体的制备及应用[J]. 畜牧兽医学报, 2019, 50(9): 1882-1887.

羊莫氏巴贝斯虫细菌人工染色体（BAC）文库的构建

Construction of Bacterial Artificial Chromosome (BAC) Library of Ovine Babesia motasi

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价