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不同类型猪小肠组织差异EST的鉴定及其SNPs分析
朱银剑,陈宏权,潘中婷,陈华,焦明慧,陈公伟,谢亚男,周梅
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(安徽农业大学动物科技学院,合肥230036;安徽农业大学动物科技学院,合肥230036; 安徽地方畜禽遗传资源保护与生物育种省级实验室,合肥230036)
摘要:
采用DDRT-PCR技术,对中国地方猪种和杜长大(大白×长白×杜洛克)猪的小肠组织进行差异EST分析。试验提取了地方猪和杜长大猪小肠组织的总mRNA,反转录成cDNA,利用3条锚定引物和6条随机引物组成18个引物对其进行PCR扩增,对特异性差异条带进行测序分析和鉴定其EST;针对EST序列特征,在不同产脂能力猪种中开展SNPs分析。结果表明,18个引物对共得到180多条EST序列,其中地方猪种个体间差异EST为6条,杜长大猪个体间差异EST为12条,地方猪与杜长大猪间差异EST为8条。选取具有代表性的3条种群间差异EST进行序列研究,结果显示,G24-300在种群之间的表达量存在明显的差异,是在杜长大猪群中高表达的EST;G01-360具有丰富的SNPs,并且其基因频率分布在不同类型猪群中存在明显的差异。
关键词:  地方猪种  小肠组织  EST  SNP  产脂能力
DOI:
基金项目:国家自然科学基金(31172180)和安徽省科技攻关项目(12010302052)共同资助。
Identification and SNPs of EST in small intestine tissue of different pig-types
ZHU Yin-jian,CHEN Hong-quan,PAN Zhong-ting,CHEN Hua,JIAO Ming-hui,CHEN Gong-wei,XIE Ya-nan,ZHOU Mei
(School of Animal Science and Technology, Anhui Agricultural University, Hefei 230036;School of Animal Science and Technology, Anhui Agricultural University, Hefei 230036; Anhui Provincial Laboratory for Local Livestock and Poultry Genetic Resource Conservation and Breeding, Hefei 230036)
Abstract:
The differential EST of small intestine tissue was investigated in Chinese native pig breeds and DLY(Duroc×Large white×Yorkshire) using DDRT-PCR. Total mRNA in small intestine tissue were extracted and reversely transcribed into cDNA. 18 primer pairs combinated by three anchor primers and six random primers were employed for differential display of reverse transcriptional polymerase chain reaction (DDRT-PCR). The PCR products were sequenced and then the differential EST and its SNPs were analyzed. The results showed that more than 180 EST sequences were obtained, including 6 inter-individual differential ESTs of the local pig breed, 12 inter-individual differential ESTs of the DLY pigs. Eight differential ESTs between the local breed and DLY pig were detected in which 3 typical differential ESTs, G24-300, G01-360 and A12-340, were selected for further research. The expression level of G24-300 in the DLY was higher than that in local pigs, while the expression levels of G01-360 and A12-340 EST didn't differ between two pig-types. G01-360 had rich SNPs, and showed the difference between native pig breed and DLY.
Key words:  pig  small intestine tissue  EST  SNP  lipogenesis ability

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