Sequence Analysis Of NGB Gene In Bactrian Camel And Its Expression In Retina

In order to analyze the sequence information of NGB gene and expression orientation of NGB protein in Bactrian Camel retina,explore the physiological funcation and which correlation with extreme natural environment,adult healthy Bactrian camel was used as the research object to obtain the NGB gene sequence using TA cloning method,the physicochemical properties and advanced structure of nucleotide sequence and encoded product were predicted and analyzed by the related software DNAStar and Predictprotein.The distribution and expression characteristic of NGB on the retina of Bactrian camel were analyzed by using the conventional paraffin section HE staining and immunohistochemical SP method.The specific findings are as follows:1.A length of 5210 bp NGB gene was obtained in Bactrian camel by TA cloning method,including 4 exons and 3 introns,the exons length were 89 bp,112bp,120 bp,135bp,and the introns length were 1505 bp,618bp and 1437 bp respectively and a Cp G island which is 257 bp.2.Amino acid sequence analysis showed that 20 kinds of standard amino acids were found in Bactrian camels NGB.In the encoded polypeptide chain,the number of different amino acids varies greatly,among which leucine(L)showed the greastest amount,occupying 13.91% of the whole.Aspartic amide(N)was accounted for 1.32% of the whole amino acid.3.The relative molecular mass of the NGB in Bactrian camels is 5.39,the Half-Life is 30 h,its instability coefficient is 56.50,which is the unstable protein,so the prediction of NGB is unstable and the protein is hydrophilic and soluble protein.The NGB secondary structure was predicted,the results showed that the ratio of α-helix 76.16%,β-folding was 0%.4.The results of HE staining of the retinal structure of Bactrian camel show that the retinal cells of Bactrian camel are distinct and have a regular 10-layer structure,which is the same as other mammals known.From the inside to the outside in order: the inner boundary membrane,optic nerve fiber layer,ganglion cell layer,inner layer,kernel layer,outer reticular layer,outer core layer,external membrane,optic rod cone layer and pigment epithelial layer.5.The results of IPP and statistics showed that there is no significant difference between MD value and negative control group in the outer core layer,the MD values of the other regions were optic nerve fiber layer,ganglion cell layer,inner reticular layer,kernel layer,outer reticular layer,photoreceptor inside,outer segment,and pigment epithelial layer.There were significant differences between MD and negative control in the above regions,and the expression of weak positive(+)was determined,and the result was that the strong positive(+ +)expression was determined by 22 compared with the MD value and the difference was significant,which was consistent with the observation results under optical microscope.6.The results may be varied among different animal species,and may be closely correlated with the function of the outer segment of the photoreceptor in the extreme environment,but it is still a problem whether it is relevant to the adaptability of the environment.