Abstract In the present study, the cloning and sequence analysis of interleukin‐6 (IL‐6) from the giant panda (Ailuropoda melanoleuca) were described. Sequencing revealed that IL‐6 complementary DNA contains a 636 base‐pair open reading frame encoding 211 amino acids. Homology analyses indicated that the identity levels of nucleotide and deduced amino acid sequences of IL‐6 between the giant panda and other species in Carnivora ranged from 90.5% to 85.6% and from 82.9% to 76.5%, respectively. Phylogenetic analyses based on nucleotide and amino acid sequences showed that the harbour seal shared a closer relationship with the giant panda than other carnivores (ursids not included). The functionally important domains and sites that have previously been recognized in other carnivores were all observed in the IL‐6 sequence of the giant panda. These findings suggest that the complementary DNA fragment cloned is indeed the IL‐6 molecule with biological activity and can be used in immunological studies of the giant panda. Keywords: Ailuropoda melanoleuca interleukin‐6complementary DNA sequencephylogeny Acknowledgements This research work was funded by a grant from Chengdu Giant Panda Breeding Research Foundation. The authors are grateful to Chengdu Research Base of Giant Panda Breeding for providing the samples of giant panda blood. Liu Xu and Bo Zeng contributed equally to this work.
Primordial germ cells (PGCs) play a crucial role in preserving poultry genetic resources and conducting transgenic research. A system for the rapid isolation of PGCs from single chicken embryonic blood was established in this paper. We found that PGCs can migrate to the lower layer of chicken embryonic fibroblasts (CEFs) through pores smaller than their diameter, while blood cells cannot, when co-cultured with CEFs of passages two to three. Based on the characteristics of PGCs, we developed a new PGC isolation method (cell culture insert/CEF adhesion method) that utilizes a 3 μm cell culture insert and CEFs of passages two to three. Using this method, approximately 700 PGCs can be isolated from the blood of a single chicken embryo at Hamburger and Hamilton (H&H) stage 17 of development. The separation rate achieved was 87.5%, with a separation purity of 95%. The separation rate of this method was 41.4% higher than the common Percoll density gradient centrifugation method and 33.6% higher than lysis with ACK buffer. PGCs isolated from embryonic blood could proliferate 37-fold within 2 weeks when cultured in a feeder-free culture system. They also continued to express the SSEA-1 and DAZL proteins and retained the ability to migrate in vivo. Overall, PGCs separated using cell culture inserts/CEF adhesion method retain their stem cell characteristics and migration ability. PGCs also exhibit good proliferation efficiency, making them suitable for subsequent transgenic experiments or genetic resource preservation.
The complete mitochondrial genome of Pallas's squirrel (Callosciurus erythraeus) from Sichuan Province was sequenced and characterized in detail. It was 16,550 bp in length and composed of 13 protein-coding genes, 22 tRNAs, 2 rRNAs and 1 control region. The mitochondrial genome of C. erythraeus presented in this report will be useful for species identification, genetic variability and clarifying the controversial taxonomic status of genus Callosciurus.
Alkaloids are important active ingredients occurring in many traditional Chinese medicines, and alkaloid glycosides are one of their existence forms. The introduction of saccharide units improves the water solubility of alkaloid glycosides thus presenting better biological activity.Because of the low content in plants, alkaloid glycosides have been not comprehensively studied. In this study, ultrahigh performance liquid chromatography-quadrupole time of flight-tandem mass spectrometry(UPLC-QTOF-MS/MS) was employed to identify and analyze the alkaloid glycosides in Coptis chinensis, Phellodendron chinense, Menispermum dauricum, Sinomenium acutum, Tinospora sagittata and Stephania tetrandra. The results showed that except Tinospora sagittata, the other five herbal medicines contained alkaloid glycosides. Furthermore, the alkaloid glycosides in each herbal medicine were identified based on UV absorption spectra, quasimolecular ion peaks in MS, fragment ions information in the MS/MS, and previous literature reports. A total of 42 alkaloid glycosides were identified. More alkaloid glycosides were identified in C. chinensis and Menispermum dauricum, and eleven in C. chinensis were potential new compounds. Furthermore, the alkaloid glycosides in the water extract of C. chinensis were coarsely se-parated by macroporous adsorption resin, purified by column chromatography with D151 cation exchange resin, ODS and MCI, combined with semi-preparative high performance liquid chromatography. Two new alkaloid glycosides were obtained, and their structures were identified by mass spectrometry and NMR data as(S)-7-hydroxy-1-(p-hydroxybenzyl)-2,2-N,N-dimethyl-1,2,3,4-tetrahydroisoquinoline-6-O-β-D-glucopyranoside and(S)-N-methyltetrahydropalmatubine-9-O-β-D-glucopyranoside, respectively. This study is of great significance for enriching the information about the chemical composition and the in-depth development of C. chinensis. Meanwhile, it can provide a reference for rapid identification and isolation of alkaloid glycosides from other Chinese herbal medicines.
The blue sheep (Pseudois nayaur nayaur) belongs to the subfamily Caprinae, and is distributed in the Qinghai-Tibet Plateau and peripheral mountains of China. In this study, the complete mitochondrial genome of P. n. nayaur was sequenced. It was determined to be of 16,700 bases. The nucleotide sequence data of 12 heavy-strand protein-coding genes of P. n. nayaur and other 29 caprine species were used for phylogenetic analysis. Phylogenetic trees were reconstructed by Bayesian and maximum likelihood methods, and it suggested that P. n. nayaur was the sister group of P. schaeferi and P. nayaur, and also indicated a close relationship of genus Pseudois with genus Capra.
With largely allopatric distribution, the black tokay and the red tokay are two distinct morphs of the subspecies Gekko gecko gecko. In consideration of their different morphological characteristics, the taxonomic status of G. g. gecko is disputed. Through detailed morphological comparison, it has been proposed that the black tokay should be elevated to species ranking, but without strong genetic evidence. In order to further investigate the taxonomic status of the tokay gecko (G. gecko), we used one mitochondrial marker (ND2) and three nuclear markers (RAG1, c-mos, and ITS2) to explore the phylogenetic and taxonomic relationship of the tokay gecko. Our results revealed a deep phylogeographical divergence in tokay gecko and at the same time provided us with the evidence of possible introgressive hybridization or/and incomplete lineage sorting between the black tokay and the red tokay. The elevation of the black tokay to species level is also supported by our results. However, due to limited sampling and genetic data, this elevation should be further corroborated by more genetic evidence.
Blue sheep, Pseudois nayaur, is endemic to the Tibetan Plateau and the surrounding mountains, which are the highest-elevation areas in the world. Classical morphological taxonomy suggests that there are two subspecies in genus Pseudois (Bovidae, Artiodactyla), namely Pseudois nayaur nayaur and Pseudois nayaur szechuanensis. However, the validity and geographic characteristics of these subspecies have never been carefully discussed and analyzed. This may be partially because previous studies have mainly focused on the vague taxonomic status of Pseudois schaeferi (dwarf blue sheep). Thus, there is an urgent need to investigate the evolutionary relationship and taxonomy system of this genus. This study enriches a previous dataset by providing a large number of new samples, based on a total of 225 samples covering almost the entire distribution of blue sheep. Molecular data from cytochrome b and the mitochondrial control region sequences were used to reconstruct the phylogeny of this species. The phylogenetic inferences show that vicariance plays an important role in diversification within this genus. In terms of molecular dating results and biogeographic analyses, the striking biogeographic pattern coincides significantly with major geophysical events. Although the results raise doubt about the present recognized distribution range of blue sheep, they have corroborated the validity of the identified subspecies in genus Pseudois. Meanwhile, these results demonstrate that the two geographically distinct populations, the Helan Mountains and Pamir Plateau populations, have been significantly differentiated from the identified subspecies, a finding that challenges the conventional taxonomy of blue sheep.
AbstractIn this study, the mitochondrial genome of Asian golden cat (Catopuma temminckii) is sequenced. The mitochondrial genome was 16,985 bp long, including 13 protein-coding genes, 22 transfer RNA (tRNA) genes, 2 ribosomal RNA (rRNA) genes, 1 control region and 1 origin of light-strand replication. The overall base composition of the mitochondrial genome was 32.76% A, 27.49 % T, 25.75 % C, and 13.99 % G. The complete mitochondrial genome of Catopuma temminckii could contribute to understanding taxonomic status and phylogenetic relationship of genus Catopuma.KeywordsAsian golden catCatopuma temminckiimitochondrial genome Declaration of interestThe authors report no conflict of interest. The authors alone are responsible for the content and writing of the paper.This research was supported by the Research Project of Chinese Ministry of Education (No.113054A).
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