为制备鸡肝脏型脂肪酸结合蛋白(L-FABP)的多克隆抗血清,并分析L-FABP的组织表达特性,利用RT-PCR扩增L-FABP基因,构建鸡L-FABP基因的GST融合蛋白表达质粒pGEX-4T/L-FABP。将重组表达质粒转化大肠杆菌BL21,IPTG诱导产生GST/L-FABP融合蛋白,用亲和层析纯化目的蛋白,将纯化的GST/L-FABP融合蛋白免疫家兔制备抗血清,并利用此抗血清分析鸡L-FABP基因的组织表达特性。诱导得到了1个40 ku(14ku L-FABP+26 ku GST)的融合蛋白,获得效价较高、特异性强的鸡L-FABP的抗血清。鸡L-FABP的组织表达特性研究结果表明,该基因在肝脏和小肠组织中有较高表达,但在心脏、脂肪、肌肉、肌胃、脾、肺和肾中没有检测到表达信号。
Lipid metabolism and adipocyte differentiation are reglulated by networking of transcription factors.It is generally known that three factors,peroxisome proliferator-activated receptorγ(PPARγ),CCAAT/element-binding proteinα(C/EBPα)and sterol regulatory element binding protein-1(SREBP1),play fundamental roles in metabolic pathways.And they are also important in adipocyte differentiation.Expressions of these factors are regulated by some compounds such as fatty acids or some steroid hormones (insulin)which is stimulated by the nutritional level.Furthermore,these factors are related to some metabolic diseases including type II diabetes and obesity.Lots of researches have focused on relationships between the factors and the genetic diseases.Different functions of factors on inducing the adipocyte differentiation are other hot spots according to previous studies.This paper summarized these studies and gave a limpid description of structures and functions of these genes.
Based on chicken' consensus map issued in 2000,17 microsatellites near 4 candidate genes such as IGF2,OBR,GDF8 and APOA1 in 4 chromosomes(chromosome 5,7,8 and 24)were chosen for polymorphism analysis and construction of linkage map.Combining the technique of PCR and the fluorescent semi-automated detection,genome scanning was performed for 440 chickens,which was derived from China Agricultural University chicken resource families within three generations.The individuals of this resource families were genotyped.The results showed that the number of alleles ranged from 4 to 14;heterozygosity(H) of markers was between 0.3116 and 0.9148.Polymorphic information content(PIC)varied from 0.2672 to 0.8679.Microsatellites along with above-mentioned 4 candidate genes doing as general markers were used to construct linkage map.The spans of 4 linkage maps constructed in the part region of chromosome 5,7,8 and 24 were 263.5,79.9,206.2 and 104.2 cM,respectively.The order of markers was consistent with that of counterpart of reported consensus map.However,The spans of linkage map were larger than that of consensus map.The constructed linkage maps laid the foundation for mapping quantitative trait loci(QTL)responsible for economically important traits in chicken.
WANG ShouzhiLI HuiLI NingGAO YuDU ZhiqiangGU ZhiliangWANG QiguiLI ZhihuiWANG Ying
