Cloning, bioinformatics and tissue expression patterns of ADD1 gene in Guizhou black goats

In order to analyze the coding sequence（CDS） information and expression patterns of the ADD1 gene in Guizhou black goats, the CDS region of the ADD1 gene in Guizhou black goats was cloned and subjected to bioinformatics analysis. The tissue expression patterns of ADD1 gene in hearts, livers, spleens, lungs, kidneys, subcutaneous fat, longissimus dorsi, and leg muscles of Guizhou black goats at 12 months, as well as the temporal expression patterns in the subcutaneous fat, longissimus dorsi, and leg muscle tissues of Guizhou black goats at 1, 3, 6, 9, and 12 months old were analyzed. The results showed that the CDS region of the ADD1 gene in Guizhou black goats was 2 304 bp long, encoding 767 amino acids. The ADD1 gene sequence had high similarity to goats and low similarity to birds. The main amino acids in the ADD1 protein were serine（Ser）, proline（Pro）, glutamic acid（Glu）, and leucine（Leu）, accounting for 9.0%, 8.7%, 8.5%, and 8.2% respectively. The molecular weight was 84.05 ku, with a molecular formula of C₃₆₈₂H₅₈₄₀N₁₀₃₆S₂₄, and a theoretical isoelectric point of 5.74. It was an unstable and hydrophilic protein, mainly located in the cell nucleus. It interacted with other proteins such as the ADD protein family（ADD2 and ADD3）, capping actin protein（CAPG）, myotrophin（MTPN）, X-box binding protein 1（XBP1）, and sterol-regulatory element-binding protein 1（SREBP1）. The secondary and tertiary structures mainly consisted of α-helices and irregular coils. The ADD1 gene was expressed in the hearts, livers, spleens, lungs, kidneys, subcutaneous fat, longissimus dorsi, and leg muscle tissues of Guizhou black goats at 12 months. The expression level was the highest in subcutaneous fat, which was significantly higher than that in other tissues（P＜0.01）, followed by the higher expression level in the hearts, longissimus dorsi, livers, and leg muscle tissues, and the lowest expression level in the spleen. The expression level of the ADD1 gene in the fat, longissimus dorsi, and leg muscle tissues of Guizhou black goats at 1 and 3 months old was low. In the fat tissue, the expression level was the highest at 6 months old, which was significantly higher than that at other ages（P＜0.01）, and the lowest level was found at 3 months old, which was significantly lower than that at other ages（P＜0.01）. In the longest back muscle, the expression level was higher at 9 and 12 months, with no significant difference between them（P＞0.05）, but both were significantly higher than that at other ages（P＜0.01）. The expression level was the lowest at 6 months old, and there was no significant difference from 1 month old（P＞0.05）, but both were significantly lower than that at other ages（P＜0.01）. In the leg muscle, the expression level was the highest at 9 months old, which was significantly higher than that at other ages（P＜0.01）, and the lowest at 12 months old, with no significant difference from 1 month old（P＞0.05）, but both were significantly lower than that at other ages（P＜0.01）. These findings indicated that the ADD1 gene was expressed in a wide range of tissues in Guizhou black goats, and the expression levels was different among different tissues.