[ Objective]The aim was to study polymorphism of mat-peptide sequence of IGF1 R gene in Baise horses and thoroughbred, which would af- ford reference for the further studies on the dwarf mechanism and molecular breedi...[ Objective]The aim was to study polymorphism of mat-peptide sequence of IGF1 R gene in Baise horses and thoroughbred, which would af- ford reference for the further studies on the dwarf mechanism and molecular breeding in horses. [Method] A total of 57 blood samples of each breed were collected and genomic DNA was extracted by the standard phenol -chloroform method. Five DNA pools of each breed were constituted and polymorphism sites were identified by sequencing PCR products. Frequencies of genotypes and alleles at these sites of each breed were checked by PCR-RFLP. [Result] Four polymorphism sites were identified in exon 2, 5 and 16, including mutations of T406C, T179 627C, G212 077A and G2.12 110A. No difference was found in the frequency of T179 627C between the Baise horses and thoroughbred. The mutation （3212 077A was only found in the thorou- ghbred, and the mutations, T406C and G212 110A, were only checked out in the Baise horses. [ Conclusion] Whether these mutations are associated with horse growth needs further studies.展开更多
Research on the identity of genes and their relationship with traits of economic importance in farm animals could assist in the selection of livestock. In this study, the polymorphisms of insulin-like growth factor 1 ...Research on the identity of genes and their relationship with traits of economic importance in farm animals could assist in the selection of livestock. In this study, the polymorphisms of insulin-like growth factor 1 （IGF1） gene in 561 goats of ten breeds were detected by polymerase chain reaction （PCR）-single strand conformation polymorphism （SSCP） and their association with litter size and birth weight in three breeds were investigated. The effects of IGF1 polymorphisms on the breeding value for litter size and birth weight were examined using least square methods. Two deletions （CA） were detected in the microsatellite and two mutations （A1637G, T1640C） were found in 5′-flanking regulatory region. No significant association between the polymorphisms in 5′-flanking region of IGF1 and birth weight was found in the three breeds of goats. In Gulin Ma goats, two polymorphisms were found to affect litter size traits. In Chuandong White goats and Guizhou White goats, no significant difference （P0.05） in litter size between goats carrying different genotypes was observed. Further evaluation and confirmation studies in more goat populations with larger sample sizes are necessary.展开更多
目的基于动物实验探讨IGF1基因慢病毒转染骨髓间充质干细胞(BMSCs)治疗骨性关节炎(OA)的作用。方法取40只SPF级雄性Wistar大鼠,10~12周龄,体重280~320 g,按照随机数字表法分为BMSCs+IGF1组、BMSCs组、模型组,每组10只,采用膝关节前交叉...目的基于动物实验探讨IGF1基因慢病毒转染骨髓间充质干细胞(BMSCs)治疗骨性关节炎(OA)的作用。方法取40只SPF级雄性Wistar大鼠,10~12周龄,体重280~320 g,按照随机数字表法分为BMSCs+IGF1组、BMSCs组、模型组,每组10只,采用膝关节前交叉韧带切断法建立OA模型,其余10只大鼠用于BMSCs的分离。BMSCs+IGF1组大鼠膝关节腔内注射0.1 ml IGF1基因慢病毒转染BMSCs(1×107个),BMSCs组大鼠膝关节腔内注射0.1 ml BMSCs(1×107个);模型组大鼠膝关节腔内注射0.1 ml生理盐水。采用酶联免疫吸附试验检测大鼠血清Ⅰ型胶原C端肽(CTX-Ⅰ)、CTX-Ⅱ、白介素(IL)-1α、IL-6水平;采用实时荧光定量聚合酶链反应(PCR)检测大鼠膝关节软骨组织中骨钙素(OC)、糖原合成酶激酶-3β(GSK-3β)、碱性磷酸酶(ALP)、Runt相关转录因子2(Runx2)的mRNA表达水平;采用Western blot检测大鼠膝关节软骨组织中β-catenin、Wnt1、GSK-3β的蛋白表达水平。结果BMSCs表面抗原CD29、CD34、CD45、CD105高表达,慢病毒转染效率(90.36±2.18)%;BMSCs+IGF1组大鼠膝关节软骨完整,无明显退化,关节面平整,其他两组均存在不同程度软骨退变,关节面不平。与模型组比较,BMSCs组血清CTX-Ⅰ、CTX-Ⅱ、IL-1α、IL-6水平,OC、GSK-3β、ALP、Runx2 mRNA表达水平,β-catenin、Wnt1、GSK-3β蛋白表达水平降低,差异有统计学意义(P<0.05);与BMSCs组比较,BMSCs+IGF1组以上指标均降低,差异有统计学意义(P<0.05)。结论IGF1基因慢病毒转染BMSCs能有效改善OA大鼠的病理损伤,能明显促进膝关节损伤修复。展开更多
文摘[ Objective]The aim was to study polymorphism of mat-peptide sequence of IGF1 R gene in Baise horses and thoroughbred, which would af- ford reference for the further studies on the dwarf mechanism and molecular breeding in horses. [Method] A total of 57 blood samples of each breed were collected and genomic DNA was extracted by the standard phenol -chloroform method. Five DNA pools of each breed were constituted and polymorphism sites were identified by sequencing PCR products. Frequencies of genotypes and alleles at these sites of each breed were checked by PCR-RFLP. [Result] Four polymorphism sites were identified in exon 2, 5 and 16, including mutations of T406C, T179 627C, G212 077A and G2.12 110A. No difference was found in the frequency of T179 627C between the Baise horses and thoroughbred. The mutation （3212 077A was only found in the thorou- ghbred, and the mutations, T406C and G212 110A, were only checked out in the Baise horses. [ Conclusion] Whether these mutations are associated with horse growth needs further studies.
基金supported by the earmarked fund for Modern Agro-Industry Technology Research System of China (nycytx-39)the Natural Science Foundation Project of CQ CSTC, Chongqing City, China (CSTC, 2009BA1066)
文摘Research on the identity of genes and their relationship with traits of economic importance in farm animals could assist in the selection of livestock. In this study, the polymorphisms of insulin-like growth factor 1 （IGF1） gene in 561 goats of ten breeds were detected by polymerase chain reaction （PCR）-single strand conformation polymorphism （SSCP） and their association with litter size and birth weight in three breeds were investigated. The effects of IGF1 polymorphisms on the breeding value for litter size and birth weight were examined using least square methods. Two deletions （CA） were detected in the microsatellite and two mutations （A1637G, T1640C） were found in 5′-flanking regulatory region. No significant association between the polymorphisms in 5′-flanking region of IGF1 and birth weight was found in the three breeds of goats. In Gulin Ma goats, two polymorphisms were found to affect litter size traits. In Chuandong White goats and Guizhou White goats, no significant difference （P0.05） in litter size between goats carrying different genotypes was observed. Further evaluation and confirmation studies in more goat populations with larger sample sizes are necessary.
文摘目的基于动物实验探讨IGF1基因慢病毒转染骨髓间充质干细胞(BMSCs)治疗骨性关节炎(OA)的作用。方法取40只SPF级雄性Wistar大鼠,10~12周龄,体重280~320 g,按照随机数字表法分为BMSCs+IGF1组、BMSCs组、模型组,每组10只,采用膝关节前交叉韧带切断法建立OA模型,其余10只大鼠用于BMSCs的分离。BMSCs+IGF1组大鼠膝关节腔内注射0.1 ml IGF1基因慢病毒转染BMSCs(1×107个),BMSCs组大鼠膝关节腔内注射0.1 ml BMSCs(1×107个);模型组大鼠膝关节腔内注射0.1 ml生理盐水。采用酶联免疫吸附试验检测大鼠血清Ⅰ型胶原C端肽(CTX-Ⅰ)、CTX-Ⅱ、白介素(IL)-1α、IL-6水平;采用实时荧光定量聚合酶链反应(PCR)检测大鼠膝关节软骨组织中骨钙素(OC)、糖原合成酶激酶-3β(GSK-3β)、碱性磷酸酶(ALP)、Runt相关转录因子2(Runx2)的mRNA表达水平;采用Western blot检测大鼠膝关节软骨组织中β-catenin、Wnt1、GSK-3β的蛋白表达水平。结果BMSCs表面抗原CD29、CD34、CD45、CD105高表达,慢病毒转染效率(90.36±2.18)%;BMSCs+IGF1组大鼠膝关节软骨完整,无明显退化,关节面平整,其他两组均存在不同程度软骨退变,关节面不平。与模型组比较,BMSCs组血清CTX-Ⅰ、CTX-Ⅱ、IL-1α、IL-6水平,OC、GSK-3β、ALP、Runx2 mRNA表达水平,β-catenin、Wnt1、GSK-3β蛋白表达水平降低,差异有统计学意义(P<0.05);与BMSCs组比较,BMSCs+IGF1组以上指标均降低,差异有统计学意义(P<0.05)。结论IGF1基因慢病毒转染BMSCs能有效改善OA大鼠的病理损伤,能明显促进膝关节损伤修复。
