转基因生物新品种培育重大专项(2008ZX08009-002, 2009ZX08009-157B)资助项目
This work were supported by grants from Domain-Specific Projects for Transgenic Biological Breeding (2008ZX08009-002, 2009ZX08009-157B)
通过3′和5′cDNA末端快速扩增(RACE)获得了猪UCP5基因全长cDNA序列,通过不同品种猪UCP5基因编码区和调控区单核苷酸多态分析,发现 -1 567 bp处存在A→T的突变,从而导致转录因子CdxA、HNF-1、Sox-5、GATA-2结合位点发生了改变.在金华×皮特兰F2代资源群中,使用PCR限制性片段长度多态性(RFLP)方法,对524头个体进行基因型测定,并与屠体性状关联分析,结果发现,AA型活体重极显著高于BB型(P < 0.01),AB型活体重显著高于BB型(0.01 < P < 0.05),AA型的后腿肌肉重极显著高于BB型(P < 0.01),AA型的后腿脂肪重显著高于BB型(0.01 < P < 0.05).同时,通过实时荧光定量PCR获得了UCP5基因在初生仔猪心、肝、脾、肺、肾、眼肌、腹脂和大脑等不同组织中基因表达的特性,结果表明,UCP5在猪的各种组织均有表达,在脑和肺中表达量最多.t检验发现,初生达兰猪眼肌中UCP5的表达量显著高于金华猪.
The cDNA of UCP5 gene was obtained by 3′ and 5′ RACE, the length was 1 611 bp. One A→T mutation was found at -1 567 bp of UCP5 gene. The mutation resulted in the change of transcript factors CdxA, HNF-1, Sox-5, GATA-2. Different genotypes were detected in 524 Jinhua× Pietrain pigs using PCR-RFLP, and the relationships between the genotypes and live weight, muscle weight of hind leg, fat weight of hind leg, average back-fat thickness, back-fat thickness in the 6th~7th rib, area of loin and leaf fat weight were analyzed. The statistical analysis showed that animals of the AA genotype had the highest live weight, which was significantly higher than that of BB genotype and obviously higher than that of AB genotype. For muscle weight of hind leg and hind leg fat weight, AA genotype was also higher than AB and BB genotype. By Real-time quantitative PCR, UCP5 expression was analyzed in heart, liver, spleen, lung, kidney, loin, abdominal adipose and cerebral tissues. The results showed that UCP5 existed in all tissues, and the expression of UCP5 was the majority in cerebral tissue and lung. The result of t-test showed that the distribution of UCP5 in loin of Dalan pigs was significantly higher than that of Jinhua.
李红霞,赵兴波,徐宁迎,姜运良,曹 梦,刘玉方,方 迟,李 宁.猪UCP5基因克隆表达特性及与屠体性状的关联分析[J].生物化学与生物物理进展,2010,37(12):1323-1330
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