巴马香猪与长白猪肌肉生长发育相关基因的研究

发布时间：2018-05-26 12:30

本文选题：巴马香猪 + 长白猪　；参考：《广西大学》2017年博士论文

【摘要】：本研究以巴马香猪为研究对象,利用RNA-seq技术和凝胶双向电泳蛋白质组学研究技术筛选30、180日龄巴马香猪背最长肌中的差异表达基因,结合肌肉代谢关键酶活性、血液生化指标、相关基因表达水平方面研究,初步揭示巴马香猪与长白猪肌肉发育及其肉质品质差异的分子机制,研究结果如下:1.巴马香猪和长白猪肉质性状、代谢相关酶活及血液生化指标的测定:(1)180日龄巴马香猪背膘厚为4.23±0.15cm,显著高于长白猪的1.47±0.21cm(p0.05);肌肉电导率为2.80±0.49ms,显著低于长白猪的4.24±0.51ms(p0.05);肌肉肉色Opto值为64.64±2.80高于长白猪的63.44±0.88,差异不显著;肌肉剪切力16.13±1.26N低于长白猪的17.89±0.96N,差异不显著;眼肌面积为13.63±0.41cm2极显著低于于长白猪的36.58±2.37 cm2(p0.01)。(2)巴马香猪和长白猪代谢关键酶活性、血液生化指标发育模式一致;0日龄时巴马香猪游离脂肪酸浓度显著高于长白猪(p0.05),LDH、TC、TG、LDL、HDL浓度显著低于长白猪(p0.05);30日龄时巴马香猪游离脂肪酸浓度、LDH、SDH、MDH、TG浓度显著高于长白猪(p0.05),TC浓度显著低于长白猪(p0.05);180日龄时游离脂肪酸浓度、LPL、SDH、MDH、TG、LDL、HDL浓度显著高于长白猪(p0.05),LDH显著低于长白猪(p0.05)。(3)巴马香猪肌肉组织LPL、LDH、MDH、SDH与肌内脂肪含量无显著性相关(p0.05)。2.巴马香猪、长白猪肌肉组织样品RNA-seq测序,结果如下:(1)30日龄和180日龄的巴马香猪、长白猪肌背最长肌肉组织样分别获得 23,051,730、21,475,968、23,000,120、18,698,878 对 clean reads,巴马香猪共拼接得到27491个转录本,19655(71.49%)个Unigene被注释到数据库中,7836(28.51%)个Unigene没有被注释,长白猪共拼接了 69686个转录本,43447(62.34%)个Unigene被注释到数据库中,26239(37.66%)个Unigene没有被注释。(2)30日龄与180日龄巴马香猪相比,有1380个差异基因,上调基因1141个,下调基因239个;2873,2713,323个基因具有一个或者多个GO注释到生物过程、细胞组成、分子功能中;206个差异基因得到KEGG Pat hway,涉及178条通路,在30日龄和180日龄巴马香猪肌肉组织中分别发现 43656、37646 个 SNP 位点。(3)30日龄与180日龄长白猪之间有351个差异基因,上调基因212个,下调基因139个;2609,1894,892个基因具有一个或者多个GO注释到生物过程、细胞组成、分子功能中;长白猪中共有258个差异基因得到了 KEGG Pathway,涉及126条通路,长白猪30日龄和180日龄肌肉组织分别发现10811、7762个SNP位点。3.巴马香猪、长白猪肌肉组织样品2-DE测定:(1)猪肌肉蛋白点的等电点(pI)值在4-7之间,分子量在25-100 kDa范围之内,总共有276个差异蛋白,149个蛋白点在30日龄时高表达,118个蛋白点在180日龄时高表达。(2)30与180日龄巴马香猪共鉴定成功了 67个差异倍数在3以上的差异蛋白,30日龄高表达的32个,180日龄高表达的35个。(3)180日龄巴马香猪与长白猪间共鉴定成了 16个差异倍数在3以上的差异蛋白,巴马香猪高表达的13个,长白猪高表达的3个。4.不同发育阶段猪背最长肌关键后选基因的表达差异分析:(1)巴马香猪和长白猪背最长肌NDUFS3、NDUFAB1、IDH3A、NU6 M、ATP5B、ATP5H、HUAMLC2B、MYL1、ADPN基因 mRNA 丰度基本一致,具有显著的年龄依赖变化,随着年龄的增加而下降(p0.05)。(2)巴马香猪背最长肌NDUFAB1、NU6M基因mRNA表达水平与肌间脂肪含量正相关,相关系数分别为:0.826(p0.05)、0.848(p0.05);A TP55H基因mRNA表达水平与肌间脂肪含量负相关,相关系数为-0.894(p0.05);HUMML2CB基因mRNA表达水平与肌肉剪切力、肌肉电导率、肌间脂肪含量负相关,相关系数分别为:-0.984(p0.01),-0.895(p0.05)、0.894(p0.05);TPI基因mRNA表达水平与肌肉剪切力、电导率极相关、肌间脂肪含量相关,相关系数分别为:0.880(p0.01)、0.852(p0.01)、0.820(p0.05);ADPN基因mRNA表达水平与肌肉剪切力相关,相关系数为:0.840(p0.05)。
[Abstract]:In this study, the differential expression genes in the longest muscle of 30180 day old Bama pig were screened by RNA-seq and gel two-dimensional electrophoresis proteomics technology, combining the activity of key enzymes in muscle metabolism, blood biochemical indexes and the water square surface of related genes, and preliminarily revealed the Bama Xiang pig and the Changbai The molecular mechanism of pig muscle development and the difference of meat quality were as follows: 1. Bama pig and long white pork quality, metabolism related enzyme activity and blood biochemical indexes: (1) the back fat thickness of Bama pig was 4.23 + 0.15cm at 180 day age, significantly higher than 1.47 + 0.21cm (P0.05) of the long white pig, and the muscle conductivity was 2.80 + 0.49ms, significantly lower than that of the muscle. The 4.24 + 0.51ms (P0.05) of the long white pig; the muscle color Opto value was 64.64 + 2.80 higher than that of the long white pig 63.44 + 0.88, the difference was not significant; the muscle shear force 16.13 + 1.26N was lower than that of the long white pig. The difference was not significant; the area of the eye muscle was 13.63 + 0.41cm2 significantly lower than that of the long white pig, 36.58 + 2.37 cm2 (P0.01). (2) Bama Xiang pig and long white pig generation. In 0 days of age, the concentration of free fatty acids in Bama pigs was significantly higher than that of long white pigs (P0.05), LDH, TC, TG, LDL and HDL were significantly lower than that of long white pigs (P0.05), and the concentration of free fatty acids in Bama pigs at 30 days of age was significantly higher than that of long white pigs at 30 days of age, and the concentration of LDH, SDH, MDH and TG was significantly higher than that of long white pigs. White pig (P0.05); free fatty acid concentration at 180 days of age, LPL, SDH, MDH, TG, LDL, HDL concentrations were significantly higher than that of long white pigs (P0.05), LDH was significantly lower than that of long white pigs (P0.05). (3) the muscle tissue of Bama Xiang pig was LPL, LDH, and muscle tissue samples were sequenced. The results were as follows: (1) 3 0 days old and 180 days old Bama pig, the longest muscle tissue like the longest muscle tissue of the long white pig obtained 23051730,21475968,23000120,18698878 to clean reads, and Bama Xiang Pigs spliced together to get 27491 transcripts, 19655 (71.49%) Unigene were annotated to the database, 7836 (28.51%) Unigene was not annotated, and the white pigs spliced 69 together. 686 transcripts, 43447 (62.34%) Unigene were annotated to the database, 26239 (37.66%) Unigene was not annotated. (2) 30 days old and 180 day old Bama pig, there were 1380 differential genes, 1141 up-regulated genes and 239 down-regulation, and 28732713323 genes have one or more GO annotated to biological processes, cell composition, molecular work In energy, 206 differentially genes were obtained by KEGG Pat hway, involving 178 pathways, and 4365637646 SNP loci were found in the muscle tissues of Bama pigs at 30 days of age and 180 days old. (3) there were 351 differential genes between 30 day old and 180 day old white pigs, up 212 and 139 down-regulation, and 26091894892 genes had one or more G. O was annotated to biological process, cell composition and molecular function; 258 different genes were obtained by KEGG Pathway, involving 126 pathways, 108117762 SNP loci.3. Bama pigs, and 2-DE determination of muscle tissue samples of long white pigs: (1) isoelectric point (pI) value of pig muscle protein points. In the range of 4-7, within the range of 25-100 kDa, there were 276 differential proteins, 149 protein points high at 30 days of age and high expression of 118 protein points at 180 days of age. (2) 30 and 180 days old Bama pigs were identified by 67 differences in 3 differences and 30 days of age high expressed. (3) the 180 day old Bama pig and the long white pig were identified as 16 differential proteins above 3, 13 high expression of Bama Xiang pig, and 3.4. expression of the longest muscle of the longest muscle in 3 different developmental stages of the long white pig. (1) the longest muscle in Bama and Changbai pigs was NDUFS3, NDUFAB1, IDH3A, NU6 M, ATP5 The mRNA abundances of B, ATP5H, HUAMLC2B, MYL1 and ADPN were basically the same, with a significant age dependence, and decreased with age (P0.05). (2) the longest muscle NDUFAB1 in Bama pig's back, and the expression level of NU6M gene mRNA was positively correlated with the intermuscular fat content, and the correlation coefficients were 0.826 (P0.05) and 0.848 (P0.05), respectively. The correlation coefficient was -0.894 (P0.05), and the level of mRNA expression of HUMML2CB gene was negatively correlated with muscle shear force, muscle conductivity, and intermuscular fat content, and the correlation coefficients were -0.984 (P0.01), -0.895 (P0.05), 0.894 (P0.05); TPI gene mRNA expression level was closely related to muscle shear force, electrical conductivity, and intermuscular fat content. The correlation coefficients were 0.880 (P0.01), 0.852 (P0.01), 0.820 (P0.05), and the ADPN gene mRNA expression level was correlated with muscle shear force, and the correlation coefficient was 0.840 (P0.05).
【学位授予单位】：广西大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：S828

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