Genetic Identification Of Myostatin Knockout Pigs And Detection Of Physiological Indexes

Myostatin(MSTN)is a negative regulator of skeletal muscle growth that has been discovered in recent years.Loss of activity can lead to excessive development of muscles in animals,increased muscle fiber diameter or increased number of myofibers,manifesting itself as an animal.Double muscle traits.Individuals with mutations in MSTN in mice,dogs,cattle,and humans all exhibit phenotypes in which skeletal muscle is overdeveloped.In this study,the unilateral mutations and double-sided mutations of MSTN gene obtained by the combination of the genome-directed editing technology and somatic cell cloning technology were tested for physiological and biochemical indicators,which provided basic data for the genetic improvement of lean-type pigs.The research content and results mainly include the following aspects:To identify the MSTN gene expression in the knockout pigs and to observe changes in their expression levels,Reverse Transcription PCR(RT-PCR)was used to detect MSTN,follistatin,Decorin),And the expression of soluble receptor ActRIIB in the longissimus dorsi and biceps femoris.After MSTN gene deletion,Follistatin,Decorin,and ActRIIB expressed differently in the MSTN+/-and MSTN-/-pigs longissimus dorsi muscles;except for a small amount of Follistatin expression in MSTN+/-biceps femoris of pigs,Decorin and ActR?B were not expressed.Follistatin,Decorin and ActRIIB were not expressed in MSTN-/-pig biceps.After testing and analyzing the blood of three pigs,wild-type WT,MSTN+/-,and MSTN-/-pigs,wild-type pigs(18.4 ± 0.55)and MSTN+/-pigs(12.25 ± 0.32)and MSTN-/-pig white blood cells were found.The total number(12 ± 0.15)was significantly different(P<0.05);however,there was no significant difference in the total number of white blood cells among MSTN knockout pigs(P>0.05);wild type pigs(22 ± 1.95),MSTN+/-Comparing the percentage of swine(15.9 ±0.92)and MSTN-/-pig neutrophils(18.7 ± 1.72)showed that there was a significant difference between the three groups(P<0.05).Urine from three groups of pigs was tested and it was found that the urobilinogen content,occult blood content,protein content and leukocyte content of the three groups of pigs all showed negligible suspected positive reactions("±");remaining bilirubin and ketones Body,nitrite,and glucose levels all showed negative reactions,indicating that the values were completely normal,and the urine pH was within the normal range(pH=7.0).In summary,although there are significant differences between some MSTN knockout pigs and wild type pigs in the above indicators,all indicators are within the normal range of healthy pigs.Therefore,it was initially determined that the indexes of the MSTN knockout pigs and the same wild type pigs were normal.