SSH文库技术结合DGGE技术筛选饲喂蜡样芽孢杆菌肉鸡脾脏免疫差异基因

Immune-Related Differentially Expressed Genes of Spleen in Broilers Fed with Probiotic Bacillus cereus Screened by SSH Library and DGGE Technique

为探究蜡样芽孢杆菌PAS38影响肉鸡免疫的分子机制,将60羽7日龄白羽肉鸡分为2组,对照组饲喂基础日粮,试验组饲喂含蜡样芽孢杆菌PAS381×106 cfu/g的基础日粮。于42日龄时从每个组随机取6只肉鸡,解剖采取脾脏并提取总RNA、反转录为cDNA。然后分别采用SSH和ACSSH技术构建脾脏差异基因文库并筛选免疫相关基因。结果显示,SSH筛选得到119个差异基因,其中上调基因63个,下调基因56个,包括9个免疫调节基因;ACSSH筛选得到5个差异基因,都为下调基因,无免疫调节基因。结果表明,饲喂蜡样芽孢杆菌能影响JCHAIN、FTH1、P2RX7等肉鸡脾脏免疫基因的表达;SSH文库明显优于ACSSH文库,ACSSH技术存在一定缺陷。

To explore the molecular mechanism of the effect of Bacillus cereus PAS38 on the immunity in broilers.607-day-old broilers were divided into two groups,the control group was fed with basal diet,and the treatment group was fed with basal diet containing Bacillus cereus PAS381×106 cfu/g.Six broilers were randomly selected from each group at the age of 42 days,spleens were taken and total RNA was extracted,then,and reversed transcription to cDNA.SSH and ACSSH techniques were used to construct splenic differential gene library and screen immune-related genes respectively.The results showed that 119 differentially expressed genes were screened by SSH,including 63 up-regulated genes and 56 down-regulated genes,and including 9 immunoregulatory genes,while five differentially expressed genes were screened by ACSSH,all of which were down-regulated genes,and no immunoregulatory genes were found.The results indicated that feeding Bacillus cereus PAS38 could affect the expression of immune genes such as JCHAIN,FTH1 and P2RX7 in the spleen of broilers,SSH library was superior to ACSSH library,and ACSSH technology was defective.