[1]史沛举,等.2型猪链球菌中国强毒株05ZYH33 ofs N-片段基因敲除株的构建[J].南京师大学报(自然科学版),2009,32(02):87-92.
　Shi Peiju,Hao Xina,Ge Junchao,et al.Deletion of the N-terminal Fragment of ofs Gene in Streptococcus Suis Serotype 2 Chinese Highly Virulent Strain 05ZYH33[J].Journal of Nanjing Normal University(Natural Science Edition),2009,32(02):87-92.
点击复制

2型猪链球菌中国强毒株05ZYH33 ofs N-片段基因敲除株的构建()
分享到：

《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

卷:
第32卷
期数:
2009年02期
页码:
87-92
栏目:
生命科学
出版日期:
2009-06-30

文章信息/Info

Title:
Deletion of the N-terminal Fragment of ofs Gene in Streptococcus Suis Serotype 2 Chinese Highly Virulent Strain 05ZYH33
作者:
史沛举1;  2;  郝喜娜1;  2;  葛俊超1;  2;  王长军2;  王晶2;  潘秀珍1;  2;  唐家琪2
1. 南京师范大学生命科学学院, 江苏南京210046
2. 南京军区军事医学研究所, 江苏南京210002
Author(s):
Shi Peiju1; 2; Hao Xina1; 2; Ge Junchao1; 2; Wang Changjun2; Wang Jing2; Pan Xiuzhen1; 2; Tang Jiaqi2
1.School of Life Sciences,Nanjing Normal University,Nanjing 210046,China
关键词:
2型猪链球菌;  of s;  基因敲除株
Keywords:
strep to coccus suis sero type 2;  ofs;  gene knock -out mutant
分类号:
R378.12
摘要:
通过生物信息学分析2型猪链球菌(Streptococcus suisserotype 2,S.suis2)中国强毒株05ZYH33的基因组,预测并发现猪链球菌血清浑浊因子(Opacity factor ofS.suis,OFS)的编码基因ofs.为了构建ofsN-末端片段ofs 37-683的基因敲除株,首先构建中间为壮观霉素抗性基因,两侧为ofs 37-683上、下游同源序列的基因敲除质粒,并对该质粒进行PCR和酶切鉴定.根据同源重组的原理,通过电转化方法筛选得到ofs 37-683基因敲除株.PCR、测序和RT- PCR 分析结果均显示ofs 37- 683已完全被壮观霉素抗性基因替代, 证实of s 37- 683基因敲除株构建成功. 该敲除株的获得为进一步研究ofs 在猪链球菌2型致病机制中的作用奠定基础.
Abstract:
The gene o f Opac ity factor o fS. suis ( ofs) w as pred icted in the g enom e o fS trep tococcus suis sero type 2 (S. su- is 2) Chinese h ighly v iru lent stra in 05ZYH 33 and its prote in structure was charac terized w ith B io informa tics too ls. To de lete theN term ina l fragment of ofs in 05ZYH 33, recom b inant gene knock ou t p lasm id w ith a Sp cr casse tte flanked w ith hom o logy arm s was constructed. The p lasm id w as confirm ed by PCR and restriction enzym e d igestion. Accord ing to the pr inciple of hom ologous recom b ina tion, the isogenic of s 37- 683 de fic ient mutant was screened through e lectroporation transform ation. PCR am plication, sequenc ing o f its produc t and RT - PCR analysis con firm ed that the target gene was replaced by Sp crcassette. Th is m utated 05ZYH 33 provided a potent too l fo r assessing the role of ofs in the S. suis 2 infection.

参考文献/References:

[ 1] Staa ts J J, Feder I, Okw um abua O, et a .l S trep tococcus suis: past and presen t[ J]. Ve t Res Commun, 1997, 21( 6): 381-407.
[ 2] Lun Z R, W ang Q P, Chen X G, et a.l S trep tococcus su is: an eme rg ing zoono tic pathogen[ J] . Lance t In fect D is, 2007, 7( 3): 201-209.
[ 3] Tang J, W ang C, Feng Y, e t a .l S treptococcal Tox ic Shock Syndrom e Caused by Strep tococcus su is Serotype 2[ J] . PLoSM ed, 2006, 3( 5) : e151.
[ 4] Sr iskandan S, Slater J D. Invasive D isease and Tox ic Shock due to Zoono tic S trep tococcus su is: An Em erg ing In fection in theEast? [ J]. P loSM ed, 2006, 3 ( 5) : e187.
[ 5] Chen C, Tang J, DongW, et a.l A G limpse o f Streptoco cca lTox ic Shock Syndrom e from Com parativeGenom ics o fS. suis 2Chinese Isolates[ J]. PLoS ONE, 2007, 2 ( 3) : e315.
[ 6] 孙雯, 潘秀珍, 王长军, 等. 猪链球菌2型烯醇化酶的分子克隆与免疫学特性[ J]. 微生物学通报, 2008, 35( 1): 15-19.
[ 7] 赵华梅, 潘秀珍, 王长军, 等. 猪链球菌2 型谷氨酸脱氢酶基因的克隆表达及酶活性测定[ J]. 中国人兽共患病学报, 2006, 22 ( 1) : 22-25.
[ 8] Baum s C G, Ka im U, Fu ldeM, et a.l Identification o f a novel v irulence dete rm inant w ith serum opac ification activ ity inS trep tococcus su is[ J]. Infection and Imm un ity, 2006, 74( 11): 6 154-6 162.
[ 9] Sm ith H E, W isse link H J, VechtU, et a .l H ighe ffic iency transform ation and gene inactivation in Streptococcus suis type 2
[ J] . M icrob io logy, 1995, 141( 1) : 181-188.
[ 10] 王花茹, 王长军, 唐家琪, 等. 猪链球菌种及其主要致病血清型多重PCR 检测方法的建立[ J]. 畜牧兽医学报,2008, 39( 6): 837-841.
[ 11] Segura M, Gottscha lk M, Oliv ierM. Encapsu lated S trep tococcus suis inh ib its activation of signa ling pa thways invo lved in
phagocyto sis[ J]. Infection Immunity, 2004, 72 ( 9): 5 322-5 330.
[ 12] Staats J J, Plattner B L, S tew art G C, e t a.l Presence o f the Strep tococcus su is su ilysin g ene and expression o fMRP and EF
correlates w ith h igh v iru lence in S trep to coccus suis type 2 iso lates[ J] . VetM icrob io ,l 1999, 70( 3 /4) : 201-211.
[ 13] Charland N, H a re l J, KobischM, et a.l Strep tococcus suis sero type 2 m utants deficien t in capsu la r express ion[ J] . M icrobi-o logy, 1998, 144( 2):325-332.
[ 14] Sm ith H E, Damm anM, van derVe lde J, et a.l Stockhofe-Zurw ieden N, Sm itsMA. Identification and character ization o f
the cps locus of S trep tococcus suis se rotype 2: the capsule protects aga inst phagocytosis and is an important v irulence factor[ J]. Infec tion Imm un ity, 1999, 67 ( 4) : 1 750-1 756.
[ 15] T imm er A M, Kr istian S A, DattaV, e t a .l Se rum opac ity factor prom otes g roup A strepto cocca l epithelia l ce ll inv asion and virulence[ J]. M olM icrob io,l 2006, 62( 1) : 15-25.

备注/Memo

备注/Memo:
基金项目: 国家自然科学基金( 30730081, 30600533, 30670105 )、江苏省自然科学基金( BK 2007013, BK2008066)、南京军区医学科技创新课题( 07Z045 )、南京军区卫生专业人才培养“ 122”工程项目.
通讯联系人: 潘秀珍, 博士, 研究员, 研究方向: 自然疫源性疾病病原等. E-m ail: panxiu zhen- 2004@ 163. com
更新日期/Last Update: 2013-04-23