| It is intramuscular fat (IMF) that plays a key factor in affecting meat quality, the genetic variation of IMF content is associated with heart type fatty acid binding protein (H-FABP), in recent years, scientists found that Small RNA (miRNA) can be combined with H-FABP which was target gene, by this way it will adjust the content of the IMF. then it will affect the quality of meat. Liaoyu white is a new breed which was cultivated by Liaoning province, in order to further improve the meat quality of the liaoyu white, the gene of H-FABP was studied in this study.There are three parts of this study:Test one:Liaoyu white, French Charolais cattle, Limousin cattle, Yanbian local cattle, The complex of cattle, Delay jiang cow were studied as objective in this study, the method is by amplificating the 3’UTR of six varieties by nesting PCR, then by means of molecular biological technology, we found a change in 3’UTR, by designing the special primers,3’UTR of H-FABP was amplified, cloned and sequenced, the length is about 230bp.Test two:The sequences are as objective which were sequencing by test one; (1) By means of molecular biological technology, H-FABP gene sequence of liaoyu white compared with the gene sequences of other varieties of cattle and looking for single nucleotide polymorphisms (SNP); Analysis results show that:There was a SNP in the 3’UTR of H-FABP gene which was in 170bp, it changed from A to G. (2) Exporing the potential miRNAs in the 3’UTR of H-FABP by online software which was Segal Lab of Computational Biology, screening results show that there were more than 400 miRNAs can be combined with H-FABP gene of Liaoyu White no matter whether it changed or not, before mutation, H-FABP can combined with bta-miR-421, bta-miR-484, bta-miR-2355-5p, bta-miR-2413, bta-miR-2461-3p, but when it has been changed, it can be combined with bta-miR-212, bta-miR-411a, bta-miR-2370-3p, bta-miR-30f, bta-miR-3604, bta-miR-2468.Test three:Liaoyu white, Limousin cattle, Yanbian local cattle are studied as objective in this part. All of these miRNA:bta-miR-421, bta-miR-484, bta-miR-2355-5p, bta-miR-2413, bta-miR-2461-3p, bta-miR-212, bta-miR-411a, bta-miR-2370-3p, bta-miR-30f, bta-miR-3604, bta-miR-2468 were designed primers by the cDNA synthesis of sangon by the first chain, then using these primers, we will begin fluorescence quantitative PCR. Experiment shows: Bta-miR-30f and bta-miR-2468 which were in the trough, which expressed lowest in the three breeds, so they may played the most important role in inhibiting affection to H-FABP, the more H-FABP gene, may be the more IMF; Bta-miR-212, bta-miR-411a, bta-miR-484, bta-miR-2355-5p, bta-miR-2413 and bta-miR-2461-3p are between the trough and the peak, they play a regulatory role in IMF, but the function is not particularly higher than bta-miR-30f and bta-miR-2468, but they are higher than bta-miR-2370-3p, bta-miR-3604 and bta-miR-421. Bta-miR-2370-3p, bta-miR-3604 and bta-miR-421 expression the highest in the three breeds, they may play the lowest important role in inhibiting affection to H-FABP, the lowest H-FABP gene, may be the lowest IMF.Conclusion:(1) The length of 3’UTR in H-FABP gene of Liaoyu White is 230bp; (2) There was a SNP in the 3’UTR of H-FABP gene which is in 170bp, it changs from A to G; There are more than 400 miRNAs can be combined with H-FABP gene of Liaoyu White no matter whether it changed or not, there are 11 miRNAs that changs when the gene of H-FABP happened mutation. (3) After by fluorescence quantitative PCR, bta-miR-2370-3p. bta-miR--3604 and bta-miR-421 expressed the most active, so they may play a negative regulation role to H-FABP gene, therefore, the lowest intramuscular fat is may deposition in the beef. On the other hand, bta-miR-30f and bta-miR-2468 are silent, so they play a negative regulation role to H-FABP gene is weak, therefore, may be the more intramuscular fat is deposition in the beef. |
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