| Hulunbuir short-tailed sheep,Baerhu sheep,Hu sheep,Han sheep and Ujimqin sheep are the main meat sheep distributed in Inner Mongolia.By resequencing the Hulunbuir sheep genome,we determined that two nucleotide site mutations in the T/Brachyury gene were responsible for the short tail of the Hulunbuir short-tailed sheep.Among them,c.G334 T was a missense mutation,and c.G333 C was a missense mutation.are synonymous mutations.c.The G334 T mutation results in the conversion of glycine to tryptophan at amino acid 112 in the T-box domain of the protein.Whether this is a sign of the difference in the T/Brachyury gene between the Hulunbuir short-tailed sheep and the Baerhu sheep,Hu sheep,Han sheep and Ujimqin sheep has not been systematically reported;nor has there been a systematic analysis of the T/Brachyury gene before and after mutation of the T/Brachyury gene.Research on structural differences.In this experiment,we first observed the differences in the tails of five types of sheep by observing the tail shape characteristics and the number of tail vertebrae,and then established a method for homozygous,heterozygous and non-mutated T/Brachyury gene mutation,and initially applied this method to the specific detection of other sheep.T/Brachyury genotype differences.This will lay the foundation for identifying the characteristics of Hulunbuir short-tailed sheep and the application of Hulunbuir mutton origin.In addition,the difference in protein secondary structure before and after mutation is studied to lay a foundation for explaining the short-tailed mechanism.According to the above background,this experiment is divided into the following parts.1.Taking the Hulun Buir sheep short-tailed sheep,Baerhu sheep,Hu sheep,Han sheep and Ujimqin sheep as the research objects,the tail morphology and anatomical structure of the tail vertebrae of five sheep were studied,the number and length of the tail vertebrae were determined,and the The genotypes at positions 333 and 334 of the second exon of the T/Brachyury gene were analyzed and compared with the tail phenotype.The results of the study found that the joints of the tail vertebrae of the Hulunbuir short-tailed sheep were smaller in number and length than those of the four types of Inner Mongolia with normal tail phenotypes of Baerhu sheep,Hu sheep,Han sheep and Ujimuqin sheep.Sheep,and deformed tail vertebrae.The average tail length and number of tail vertebrae of 20 Hulunbuir short-tailed sheep,Baerhu sheep,Hu sheep,Han sheep and Ujimqin sheep were 13.6cm,20.2cm,16.1cm,15.3cm and 32.6cm,respectively.The average number of coccygeal vertebrae were 11.8,15,14.7,13,and 16,respectively.2.In the experiment,the In-Fusion method was used to extract the c DNA amplification products of 16-day short-tailed sheep and long-tailed sheep embryos after reverse transcription as the target fragment,and the His-BRACHYURY expression vectors of short-tailed sheep and long-tailed sheep were constructed.Synthesis of BRACYURY recombinant protein,through circular dichroism analysis and statistics found that the short-tailed sheep and big-tailed sheep BRACYURY protein CD patterns in the wavelength of 190～260nm have certain differences,and the proportions of some structural types are also different.3.Taqman probe genotyping technology for T/Brachyury gene c.G333 C and c.G334 T of sheep,which are common species in Inner Mongolia Autonomous Region,was successfully established.The detected results were fed back with the T/Brachyury gene DNA sequencing results.Conclusion: Only the 333 rd and 334 th genotypes in the second exon of the Hulunbuir short-tailed sheep T/Brachyury are the two genetic genotypes of "CT/CT" homozygous and "CT/GG" heterozygous phenomenon.,Hu sheep,Han sheep,Ujimqin sheep T/Brachyury do not have this feature;established Taqman probe technology can be used for this genotyping.The mutation of T/Brachyury gene leads to obvious changes in the secondary structure of the expressed protein at the translational level. |
PDF Full Text Request