| 主办单位: |
| 中国科学院遗传与发育生物学研究所 |
| 中国遗传学会 |
| 编辑单位: |
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《遗传学报》 编辑部
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| 主 编: 薛勇彪 |
| 主 任: 张颖 |
编委阵容 |
| 审稿专家 |
| 国内刊号： |
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CN 11-5450/R
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| 国际刊号： |
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ISSN 1673-8527
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| 邮发代号： 2-819
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遗传学报 2004.3 Vol 31, No.3 258-264
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| 纯种与杂种鸡之间肝脏组织基因差异表达及其 与肉用性状杂种优势的关系 |
| Gene Differential Expression of Liver Tissues in Crossbred Versus Purebred Chicken and Their Relationship with Heterosis of Meat Trait |
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| 王 栋，张 沅①，孙东晓，俞 英，徐桂云，李俊英 |
| 中国农业大学动物科技学院遗传育种系，北京 100094 |
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| Key Words： 鸡;杂种优势;mRNA差异显示;基因差异表达模式 |
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Abstract
以白洛克肉鸡(EE)、中国丝羽乌骨鸡(CC)、农大褐（DD）和白来航（AA）4个纯种鸡为材料，进行4×4完全双列杂交，共得到16种杂交组合。应用mRNA差异显示技术（DDRT-PCR）检测了8周龄纯种和杂种鸡之间肝脏组织基因的差异表达。结果表明，在纯种和杂种间共有8种15类基因差异表达模式，杂种和纯种之间基因表达存在明显的差异。对各种基因差异表达模式与10个肉用性状的杂种优势率进行相关分析发现，表达一致型P8（t1111）与肉用性状的杂种优势率相关不显著（P＞0.05），这说明杂种优势的形成与某些基因的差异表达有关；正交或反交特异表达型P4（t0100、t0010）与8周龄个体重、腿肌重、半净膛重、全净膛重相关显著（P＜0.05），与胸肌重相关极显著（P＜0.01）；单亲特异表达型P1（t1000、t0001）与腹脂重相关显著（P＜0.05），与体斜长相关极显著（P＜0.01）；双亲特异表达型P7（t1001）与腿肌重、翅重、半净膛重、肌间脂宽的杂种优势率相关显著（P＜0.05）；正交或反交单亲表达一致型P2（t1100、t0011、t1010、t0101）与肌间脂宽的杂种优势率相关显著（P＜0.05）；单亲表达一致型P5（t1110、t0111）胫骨长的杂种优势率相关显著（P＜0.05)。 |
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Abstract
The concept of heterosis has already been put forward for a century.The hypothesis of Dominance,Superdominace and Epistasis has also been brought forward to explain the phenomenon of heterosis.As we know,there is spatio-temporal speciality about the expression of gene and only expressed genes contribute to the formation of heterosis.So the study on heterosis in expression level becomes more meaningful.A lot of studies on heterosis in this level have been done in plants,but there is no such study carried on animals in this area.In this study,the technique of mRNA Reverse Transcription Differential Display was used to research the heterosis molecular mechanism of animal.In order to expound the molecular genetic mechanism of animals heterosis,the 4×4 completely diallele cross experiment of 4 purebreds chicken was conducted among White Polymouth Rock (EE),Chinese Silk Chicken (CC),CAU Brown (DD) and White Leghorn (AA).The chicken of 16 cross combinations were reared to 8 weeks old,then 30 chicken in each combination were selected randomly and slaughtered.The traits of body weight of 8 weeks,wing weight,eviscerated weight,eviscerated weight with giblet,breast muscle yield,leg muscle yield,body length,abdomen fat weight,interamuscular fat width,tibia length were measured,and in which 8 individuals in each combination were selected randomly to collect the liver tissue samples,which were stored in liquid nitrogen or at －80℃ to be used for total RNA (TRNA) extracting.After the total RNA(TRNA) was extracted,16 TRNA pools were formed in the same quantitative according to the concentration of 8 individual TRNA.They were reversely transcribed with three anchor primers H-T11A,H-T11G and H-T11C.Then the reverse transcription PCR for each transcript product was done in two repeats at the same time with the same anchor primers and 8 random primers.The polyacrylamide gel electrophoresis of each PCR product was run in Bio-Rad Power 3 000 temperature control system.After electrophoresis,the gel was stained by AgNO3 according to the stain method described by Echt et al.The differential display bands in the polyacrylamide gel were counted.The band displayed is counted as 1 whereas no band is counted as 0 and only expression bands reproducible in two repeats were statistically analyzed.The correlation analysis between heterosis percentage and gene expression patterns was done with statistic analysis software (SAS) package.The statistic results indicated that among 690 total numbers of bands,the percentage of differential expression bands reproducible (457) is 66.23%.Eight kinds of gene differential expression patterns were found and listed as follow:1):Band presents only in one purebred (P1);2):Band in one crossbred and its corresponding paternal purebred;or Band in one crossbred and its corresponding maternal purebred (P2);3):Band in purebreds and one crossbred (P3);4):Band only in one crossbred (P4);5):Bands in both crossbreds and one purebred (P5);6):Bands only in both crossbreds (P6);7):Bands only in purebreds (P7);8):Bands both in purebreds and crossbreds P8.The differential expression of gene between purebred and crossbred chicken was detected for the first time.The proportion of each pattern in each kind of purebred combination is different.The percentage of P8 (75.34%) is the highest.The total percentage of differential expression patterns (24.66%) showed that the gene differential expression exists as a matter of fact.Among all the gene differential expression patterns, the percentage of P3 is the highest whereas the percentage of P7,P6 and P4 is very low,it indicated that different genes may have different expression patterns in purebreds and crossbreds.The results are similar to the study results on plants,which indicates that the gene differential expression between purebred and crossbred exists universally in biology.The correlation between gene expression patterns and heterosis percentage was studied,but correlation between P8 and the heterosis percentage is not significant (P＞0.05),it indicates that some patterns of gene differential expression may be the molecular genetic basic of heterosis.Among all the gene differential expression patterns,each pattern affects the expression of meat trait in different manner.There is significantly negative correlation between P4 and heterosis percentage of body weight of 8 weeks,breast muscle yield,leg muscle yield,eviscerated weight with giblet and eviscerated weight (P＜0.05);P1 is of significantly negative correlation with heterosis percentage of abdomen fat weight (P＜0.05) and of very significantly negative correlation with heterosis percentage of body lengtht (P＜0.01)；The negative correlation between P2 and heterosis percentage of interamuscular fat width is significant (P＜0.05)； The positive correlation between P7 and heterosis percentage of leg muscle yield,wing weight,eviscerated weight with giblet and interamuscular fat width is significant (P＜0.05);The positive correlation between P5 and heterosis percentage of tibia length (P＜0.05) is significant.These results show that these 5 kinds of patterns play important role in heterosis forming of meat trait.P1 and P7 show that expressed gene in purebreds is depressed;P4 indicates that new gene expression occurs in crossbreds;P5 reveals that expressed gene only in one purebred express in all crossbreds.All genes of crossbreds come from purebred,which are not only the simple adding of these purebred genes,giving birth to unknown interaction between these genes coming from different purebreds,then leading to differential expression of genes.These gene differential expressions maybe form the heterosis of meat trait. |
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中国科学院遗传与发育生物学研究所
