cDNA cloning and sequence analysis of ribosomal protein S7 gene from giant panda / 第三军医大学学报

Objective To explore the structure of ribosomal protein S7 ( RPS7) gene of giant panda ( Ailuropoda melanoleuca) and investigate its homologies with other already reported sequences,including Homo sapiens,Mus musculus,Rattus norvegicus and Bos taurus. Methods The cDNA of RPS7 was cloned from the giant panda by RT-PCR. The sequence data were analyzed by GenScan software. Blast 2. 1 was used to study the homology of the obtained RPS7 sequence with the gene sequences of other species; Open reading frame ( ORF) of the DNA sequence was searched using ORF finder software; Protein structure of the RPS7 sequence cloned was deduced using Predict Protein software. Results The full length of the sequence fragment was 589 bp containing an ORF of 585 bp. The deduced protein sequence showed that the protein was composed of 194 ami- no acids and its estimated molecular weight was 22. 126 85 ?103 with a pI of 10. 09. There were 7 different pat- terns of functional sites: one N-glycosylation site; two cAMP and cGMP-dependent kinase phosphorylation sites; four casein kinase C phosphorylation sites; one casein kinase Ⅱ phosphorylation site; two N-myristoylation sites; two amidation sites and one ribosomal protein S7e signature site in the RPS7 protein. Further analysis indicated that the sequence of RPS7 and the protein encoded were highly homologous to some mammals reported.Conclusion The complete coding sequence of RPS7 gene has been cloned through RT-PCR successfully, which is the first report on the RPS7 gene from the giant panda.

Detection of the creatine kinase activity and CK isozyme activity of black bear / 第三军医大学学报

Objective To detect the creatin kinase (CK) and its isozeyme (CK-MB) activity in serum of the healthy black bear (Ursus thibetanus) reared in pens to establish reference standard of biochemistry for CK and CK-MB activities of black bears as well as to offer scientific basis for prevention and cure of the black bear disease. Methods The CK activity and CK-MB activity were measured using spectrophotometer based on the principle of enzymatic reaction rate. All data were used to calculate the mean value, variation coefficient and the confidential interval of expected value of enzymatic activity. Results Under 30 ℃, of the male black bears, the average CK activity is 162.70 U/L, the variation coefficient 11. 90%, the confidential interval of expected value 148.90 to 176.50 U/L, and the average CK-MB activity is 17.4 U/L, the variation coefficient 17%, the confidential interval of expected value 15.26 to 19.54 U/L. Of the female black bears, the average CK activity is 149.3 U/L, the variation coefficient 8%, the confidential interval of expected value 140.68 to 157. 92 U/L, and the average CK-MB active is 15.8 U/L, the variation coefficient 18.6%, the confidential interval of expected value 13.70 to 17.90 U/L. Conclusion The data we obtained can serve as biochemical indexes for the healthy black bears.

Sequence analysis of mtDNA NADH-ubiquinone oxidoreductase chain 4 gene of Asian black bear Sichuan subspecies / 第三军医大学学报

Objective To investigate the sequence characteristic of black bear's mitochondrial NADH-ubiquinone oxidoreductase chain 4 gene(ND4).Methods Primers were designed according to certain species' NADH-ubiquinone oxidoreductase ND4 in which the DNA sequence had been reported.The DNA sequence was amplified by PCR from the Asian black bear Sichuan subspecies's(Ursus thibetanus mupinensis)hair-follicle tissue total DNA and were analyzed with its amino acid sequence.Results The ND4 gene sequence length was 1 402 bp and contained a 1 377-nucleotide open reading frame encoding 459 amino acid residues.The amino acid sequence's pI was 9.37 and weight was 51.5?10 3.The DNA sequence and corresponding amino acid sequence of black bear's ND4 gene had the high similarity with those of other mammalian animals' ND4 gene,and the similarity was 89% to 91% and 94% to 95%.The result of phylogenetic tree was basically consistent with the traditional species system phylogenetic relations.Topology prediction showed the Asian black bear Sichuan subspecies's ND4 proteins contained N-glycosylation site,kinase C phosphorylation sites,casein kinase Ⅱ phosphorylation site,N-myristoylation site and leucine-rich region.In addition,black bear's ND4 protein contained leucine zipper pattern and Leucine-rich region.Conclusion The mammalia animal's ND4 gene has the high conservative nature.The pretein coded by ND4 gene has high uniformity in the function.

Cloning and sequence analysis of mtDNA cytochrome c oxidase subunitⅠ gene of the Asian Black Bear Sichuan Subspecies / 第三军医大学学报

Objective To explore the features of the cytochrome c oxidase subunit Ⅰ (COXⅠ) of Asian black bear Sichuan subspecies. Methods Primers were designed according published gene sequences of other bear species. A DNA sequence was amplified by PCR from the bear Sichuan subspecies’s hair-follicle tissue total DNA, then the PCR product was sequenced. The results were analyzed and a forecast analysis was carried on. Results The amplified COXⅠ gene of Sichuan black bear was 1 605 bp and contained a 1545-nucleotide open reading frame encoding 514 amino acid residues. The overall base composition of COXⅠ gene is 26.9 %, 23.1%, 18.3 %, and 31.8 % respectively to A, C, G, and T. The start codon of the COXⅠ is ATG, while the termination codon TAA. The protein’s pI is 6.29 and weight is 57.2?10 3 . The similarity analysis and phylogenetic tree analysis indicated that Sichuan black bear has closer relationship to American black bear, but not to brown bear and polar bear. Among dogs, caws and rats, it is closer to the former one. Conclusion There are high similarities in the DNA sequence, protein amino acid sequence of Asian black bear Sichuan subspecies with other bear species, indicating a high conservativity of COXⅠ in these involved species.