A/J小鼠感染猪链球菌2型后脾脏消减cDNA文库的构建及相关差异基因表达分析

Construction of a suppression subtraction hybridization cDNA library to screen differentially expressed genes and functional analysis from the spleen of A/J mouse strains with infected Streptococcus suis type 2

为了分离与猪链球菌2型感染密切相关的基因,利用抑制性消减杂交(SSH)技术构建了8周龄A/J小鼠感染猪链球菌2型后的脾脏消减cDNA文库,并对其中169个阳性克隆进行测序,去除冗余的cDNA序列载体并聚类拼接后共获得110条差异表达序列标签(EST)。利用GenBank的BLAST软件分析比较核酸和蛋白质同源性,发现共有36个基因与这些EST具有高度的同源性,它们分别参与细胞成分、免疫、信号转导、凋亡、转录等重要功能。这些差异表达基因主要有ATP/GTP结合蛋白1(Agtpbp1)、天冬氨酸-谷氨酸-丙氨酸-天冬氨酸盒多肽5、核蛋白p68、ATP-依赖RNA解旋酶、真核生物转录因子2亚基(Eif2s2)、Na+/K+-ATP酶转运β3多肽、溶菌酶1、溶菌酶2等基因。结论:这些差异表达基因在猪链球菌2型感染过程中可能发挥重要功能。

This study aimed to screen the differentially expressed genes related to the infection of Streptococcus suis type 2（SS2）.The cDNA library were constructed from the 8-week-old age spleen of A/J mouse strains infected to SS2 by suppression subtraction hybridization（SSH）.A total of 169 positive clones were sequenced.After dislodging,clustering and splicing the redundant cDNA sequences,a total of 110 differentially expressed sequence tags（EST）were obtained in the whole subtractive cDNA library.We used the software BLAST in NCBI of GenBank to do the homology comparisons of nucleic acid and protein.The results showed that 36 differentially expressed genes had highly homologous with EST database.Those gene fragments involved the cell components,immunity,cell signal transduction,apoptosis,transcriptional factor genes and so on.The expression of ATP/GTP binding protein 1（Agtpbp1）,Asp-Glu-Ala-Asp box polypeptide 5,nucleoprotein-p68,ATP-dependent RNA helicase,eukaryotic translation initiation factor 2 subunit 2（Eif2s2）,ATPase/Na＋/K＋ transporting β3 polypeptide（Atp1β3）,lysozyme 1（Lyz1）and lysozyme 2（Lyz2）were significantly different after the infection of SS2 in the spleen of A/J mouse.The results indicated that those genes played an important part in the infection of SS2.