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Girardia tigrina, a freshwater planarian species native to America and introduced to other continents, has been usually used as model organism in many research fields of biology. In this study, we determined the complete mitochondrial genome of G. tigrina using next-generation sequencing (NGS). The complete mitogenome was 15,938 bp in length, with 36 genes, including 12 protein-coding genes, 2 ribosomal RNAs and 22 transfer RNAs, and ATP8 was absent in the mitogenome of G. tigrina as in the mitogenomes of some other flatworms. The maximum-likelihood phylogenetic tree suggested that G. tigrina was closely related to genus Dugesia in the clade of Tricladida.

Our previous study based on the transcriptome profiling indicated that a fragment of α-crystallin type heat shock protein (α-Hsp) gene was one of the numerous cDNA sequences expressed differentially at various stages of head regeneration in Hydra vulgaris. To further investigate the role that which α-Hsp plays during hydra regeneration, a full-length cDNA of α-Hsp gene of H. vulgaris was isolated by the rapid amplification of cDNA ends (RACE) technique. The full-length cDNA of α-Hsp gene was 1156 bp, containing a 765 bp open-reading frame (ORF), which encodes a polypeptide of 254 amino acid residues with a molecular weight of 29.27 kDa. Further, the ORF was subcloned into the plasmid pET-42a(+), and the recombinant plasmid pET-42a(+)-α- Hsp was transformed to Escherichia coli BL21(DE3), then the fusion protein GST-α-Hsp was expressed mainly in the form of a soluble molecule after induction by isopropyl-β-d-thiogalactopyranoside. In addition, BALB/Cmice were immunized with the fusion protein to prepare the polyclonal antiserum which was used as the primary antibody for whole-mount immunohistochemical assay. The results from the immunohistochemical assay showed that α-Hsp had expressedmainly at the wound site and nearby area of hydra after decapitation operation, and both quantitative real-time polymerase chain reaction (qPCR) analysis and immunohistochemical assay revealed that the expression level of α-Hsp increased gradually during the early period of hydra regeneration, then reached a peak at 24 h after decapitation operation, while decreased during the late regeneration period. Moreover, it indicated an important role of α-Hsp gene in hydra head regeneration that RNA interference (RNAi)-mediated α-Hsp silencing led to the obvious delay of the regeneration of head structures in H. vulgaris. In conclusion, our results gave the hint that α-Hsp could be related to wound healing and tissue remodelling at early regeneration stages, and may lay the foundation for further studies about the physiological function and role of α-Hsp during hydra regeneration.

Our previous study based on the transcriptome profiling indicated that a fragment of α-crystallin type heat shock protein (α-Hsp) gene was one of the numerous cDNA sequences expressed differentially at various stages of head regeneration in

A rapid and valid reversed-phase high performance liquid chromatography (RP-HPLC) method for determination of recombinant human interferon omega (rhIFNomega) in the yeast Pichia pastoris fermentation broth was developed. The method is based on the hydrophobicity of rhIFNomega followed by RP-HPLC separation with UV detection. The chromatography analysis was performed on EC 250/4 NUCLEOSIL 300-5 C18 (250 mm x 4 mm i.d., 300 A, with a particle size of 5 microm) column. The compositions of the mobile phase A and B were 999:1 (v/v) water/TFA and 999:1 (v/v) acetonitrile/TFA at a flow rate of 1.0 ml min(-1). Detection was done by spectrophotometry at 280 nm and the column temperature was 30+/-1 degrees C. Calibration curve was linear (r=0.9986, n=7) in the range of 0.074-0.555 mg ml(-1) for rhIFNomega and the regression equation was y=2.02 x 10(6)x-1.27 x 10(5). Limit of detection for rhIFNomega was 0.053 mg ml(-1). The values of R.S.D. (%) of intra-day and inter-day precision were

The complete mitochondrial genome of Oxyopes sertatus is a circular molecule of 14,442 bp in length, containing 13 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNAs, and a control region. The A + T content of the overall base composition of H-strand is 75.9% (T: 42.9%; C: 8.2%; A: 33.0%; G: 15.9%). COII, COIII and ND4 genes begin with TTG as start codon; ATP6, COI, ND1 and ND5 genes begin with ATA as start codon, ATP8, Cyt b, ND2 and ND3 genes begin with ATT as start codon, ND6 gene begins with GTG as start codon, while ND4L gene start with a typical ATG initiation codon. ND2 gene is terminated with TAG as stop codon, Cyt b and ND5 end with TA, COI, ND1 and ND4L end with T, ATP6, ATP8, COII, COIII, ND3, ND4 and ND6 end with TAA.

The complete mitochondrial genome of Selenops bursarius is a circular molecule of 14,272 bp in length, containing 13 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNAs and a control region. The A + T content of the overall base composition of H-strand is 74.4% (T: 41.8%; C: 8.7%; A: 32.6%; G: 16.9%). COI gene begins with TTA as start codon; COII gene begins with GTG as start codon; ATP6 and ATP8 genes begin with ATA as start codon; COIII and ND4 genes begin with TTG as start codon; ND1 and ND5 genes begin with ATC as start codon, while other five protein-coding genes start with ATT. ATP6, ATP8, COII, COIII, ND2, ND3, ND4 and ND5 genes are terminated with TAA as stop codon, ND1 ends with TAG, COI ends with TG, Cyt b, ND4L and ND6 end with T.

Juema pig is a kind of rare and special pig which is well adapted to high altitude, cold climate and harsh natural environment. The complete mitochondrial genome of Juema pig Sus scrofa is a circular molecule of 16 532 bp in length, containing 13 protein-coding genes, two ribosomal RNAs, 22 transfer RNAs, and a control region. The A + T content of the overall base composition of H-strand is 60.7% (T: 26.2%; C: 26.0%; A: 34.5%; G: 13.3%). ND4L gene begins with GTG as start codon, ND2, ND3, and ND5 genes begin with ATA as a start codon, and other nine protein-coding genes start with ATG. Cyt b gene is terminated with AGA as stop codon, ND1 and ND2 genes are terminated with TAG as stop codon, COII, COIII, ND3, and ND4 end with T, while ATP6, ATP8, COI, ND4L, ND5, and ND6 end with TAA. In addition, the phylogenetic relationships from neighbor-joining analyses based on the 13 concatenated PCGs indicated (Tylopoda (Suina (Ruminantia (Hippopotamidae, Cetacea)))).

The stretch spider Tetragnatha maxillosa (Araneae: Tetragnathidae) is found all over the world. In the present study, we investigated the complete mitochondrial genome of T. maxillosa and the mitogenome is a circular molecule of 14 414 bp in length, consists of 13 protein-coding genes, two ribosomal RNAs, 22 transfer RNAs, and a control region. The A + T content of the overall base composition of H-strand is 74.5% (T: 40.4%; C: 9.6%; A: 34.1%; G: 15.9%). COI gene begins with TTT as start codon, COII and COIII genes begin with TTG as start codon, ATP8, Cyt b, ND2, and ND4L genes begin with ATT as start codon, and other six protein-coding genes start with ATA. ATP6, ATP8, COI, COIII, ND1, ND3, and ND6 genes are terminated with TAA as stop codon, Cyt b, ND2, ND4, ND4L, and ND5 end with T, and COII ends with TAG. In addition, the phylogenetic relationships from neighbor-joining analyses based on the 13 concatenated PCGs indicated (Tetragnatha (Nephila (Argiope (Araneus, Neoscona)))).

The complete mitochondrial genome of Hydra vulgaris (Hydroida: Hydridae) is composed of two linear DNA molecules. The mitochondrial DNA (mtDNA) molecule 1 is 8010 bp long and contains six protein-coding genes, large subunit rRNA, methionine and tryptophan tRNAs, two pseudogenes consisting respectively of a partial copy of COI, and terminal sequences at two ends of the linear mtDNA, while the mtDNA molecule 2 is 7576 bp long and contains seven protein-coding genes, small subunit rRNA, methionine tRNA, a pseudogene consisting of a partial copy of COI and terminal sequences at two ends of the linear mtDNA. COI gene begins with GTG as start codon, whereas other 12 protein-coding genes start with a typical ATG initiation codon. In addition, all protein-coding genes are terminated with TAA as stop codon.

The complete mitochondrial genome of Chinese green hydra, Hydra sinensis (Hydroida: Hydridae) is a linear molecule of 16,189 bp in length, containing 13 protein-coding genes, small and large subunit ribosomal RNAs, methionine and tryptophan transfer RNAs, a pseudogene consisting of a partial copy of COI and terminal sequences at two ends of the linear mitochondrial DNA. The A + T content of the overall base composition of H-strand is 77.2% (T: 41.7%; C: 10.9%; A: 35.5%; and G: 11.9%). COI and ND1 genes begin with GTG as start codon, while other 11 protein-coding genes start with a typical ATG initiation codon. COII, ATP8, ATP6, COIII, ND5, ND6, ND3, ND1, ND4 and COI genes are terminated with TAA as stop codon, ND4L ends with TAG, ND2 ends with TA and Cyt b ends with T.

The amino group PEGylation of rhIFNomega with monomethoxy polyethylene glycol succinimidyl succinate (mPEG-SS, 20 000) was investigated, and the modified mixture was separated and purified by ion exchange chromatography and gel filtration chromatography. Under the optimized purification conditions, the average content ofmono PEG-rhIFNomega in the collect liquid reached 182 microg x mL(-1). The average purified yield of mono PEG-rhIFNomega exceed to 22%, and the purity of mono PEG-rhIFNomega was greater than 98% by SDS-PAGE and RP-HPLC. Relative molecular mass of mono PEG-rhIFNomega was 43 790 detected by MALDI-TOF MS. The apparent molecular mass measured by SDS-PAGE was about 60 810. The purified PEG-rhIFNomega has the characteristics of typical PEGylated protein. Activity reservation rate of mono PEG-rhIFNomega was 15.0%, while the antigenicity decreased by at least 64 folds. In addition, the acid stability, thermal stability and stability in serum and trypsin solution of mono PEG-rhIFNomega were markedly better than those of the rhIFNomega. The pharmacological properties of mono PEG-rhIFNomega were significantly improved. The prepared PEG-rhIFNomega might be developed to a novel safe and long-acting interferon.

The complete mitochondrial genome of Sinanodonta woodiana is a circular molecule of 16256 bp in length, containing 14 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNAs, and 2 control regions. The A + T content of the overall base composition of H-strand is 65.9% (T: 28.0%; C: 22.3%; A: 37.9%; G: 11.8%). F ORF (Female-specific open reading frame) begins with ATA, Cyt b begins with ATC, ATP6, ATP8, COII, COIII, ND1, ND2, ND3 and ND5 begin with ATG, ND4L begins with GTG, COI begins with TTG, and other two protein-coding genes begin with ATT as start codon. COII, COIII, F ORF, ND1, ND3, ND5 and ND6 genes are terminated with TAA as stop codon, ATP6, ATP8, COI, Cyt b, ND4 and ND4L end with TAG, and ND2 gene ends with T.

The complete mitochondrial genome of Pardosa laura is a circular molecule of 14,513 bp in length, containing 13 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNAs, and a control region. The A + T content of the overall base composition of H-strand is 77.4% (T: 42.9%; C: 8.1%; A: 34.5%; G: 14.5%). ATP6, COII and COIII genes begin with TTG as start codon; ND4 and ND6 genes begin with ATA as start codon, while other eight protein-coding genes start with ATT. COII, COIII, ND2 and ND6 genes are terminated with TAG as stop codon, COI and ND4L end with T, ND4 ends with TA, ATP6, ATP8, Cyt b, ND1, ND3and ND5 end with TAA.

The complete mitochondrial genome of Argiope amoena is a circular molecule of 14,121 bp in length, contains 13 protein-coding genes, 2 ribosomal RNAs, 21 transfer RNAs genes and a control region. The A + T content of the overall base composition of H-strand is 72.1% (T: 38.2%; C: 10.6%; A: 33.9%; G: 17.3%). ND1, ND4, ND6 and ATP6 begin with ATA as start codon, ND4L begins with ATG, ATP8, Cyt b, ND2 and ND3 genes begin with ATT, and the other two protein-coding genes begin with TTG. ATP6, ATP8, COI, COII, COIII, Cyt b, ND1, ND2, ND3, ND4L and ND5 genes are terminated with TAA as stop codon, ND6 ends with TAG, and ND4 ends with T.

The complete mitochondrial genome of Plexippus paykulli is a circular molecule of 14,316 bp in length, containing 13 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNAs and a control region. The A + T content of the overall base composition of H-strand is 73.5% (T: 40.6%; C: 9.0%; A: 32.9%; G: 17.5%). ATP6, Cyt b, ND2, ND5 and ND4 genes begin with ATA as start codon; ATP8, ND1, ND3 and ND4L genes begin with ATT as start codon; COII and COIII genes begin with TTG as start codon; COI gene begins with AGA as start codon, while ND6 gene start with a typical ATG initiation codon. ATP6, COI, COII, COIII and ND3 genes are terminated with TAA as stop codon, ND6 ends with TA, ATP8, ND1 and ND2 end with TAG, Cyt b, ND5, ND4 and ND4L end with T.

The complete mitochondrial genome of Pholcus sp. is a circular molecule of 14,279 bp in length, containing 13 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNAs, and a control region. The A + T content of the overall base composition of H-strand is 65.8% (T: 39.1%; C: 10.7%; A: 26.7%; G: 23.5%). COII and ND2 genes begin with GTG as start codon, COI, ND4L and ND5 genes begin with ATA as start codon, ATP8, ND1 and ND3 genes begins with ATT as start codon, while other five protein-coding genes start with a typical ATG initiation codon. ATP6, ATP8, COIII, ND2, ND4, ND4L and ND6 genes are terminated with TAA as stop codon, COI, Cyt b, ND1 and ND3 end with TAG, while COII and ND5 end with T.

The complete mitochondrial genome of Nephila clavata is a circular molecule of 14,433 bp in length, containing 13 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNAs, and a control region. The A + T content of the overall base composition of H-strand is 76.1% (T: 40.0%; C: 9.2%; A: 36.1%; G: 14.7%). COI gene begins with TTA as start codon, COII gene begins with TTT as start codon, COIII gene begins with TTG as start codon, ND3, ND4L, ND6 and ATP8 genes begin with ATT as start codon, while other six protein-coding genes start with ATA as initiation codon. ND3, ND4, ND4L, ND5, COI, COII, ATP6 and ATP8 genes are terminated with TAA as stop codon, ND1, ND2, ND6 and Cyt b end with TAG, and COIII ends with TGA.

The complete mitochondrial genome of Argiope bruennichi is a circular molecule of 14,063 bp in length, containing 13 protein-coding genes, 2 ribosomal RNAs, 21 transfer RNAs, and a control region. The A + T content of the overall base composition of H-strand is 73.4% (T: 38.0%; C: 9.8%; A: 35.4%; G: 16.8%). ND6 begins with ATG, ND4 and ATP6 begin with ATA, ND5 and COI begin with TTA, ND1, COII and COIII begin with TTG, and other five protein-coding genes begin with TTG as start codon. ND5, ND4L, ND6, COI, COII, COIII, ATP6 and ATP8 genes are terminated with TAA as stop codon, Cyt b, ND1 and ND2 end with TAG, and other two protein-coding genes end with T.

RT-PCR was conducted with one degenerate primer designed according to repetitive regions' amino acid sequence of major ampullate spidroin (MaSp) in spiders and adaptor primer in the SMART cDNA Library Construction Kit. By cloning and sequencing of amplified products, one cDNA clone (GenBank Accession No. AY365017) of Argiope amoena MaSp gene was obtained. The deduced amino acid sequence can be distinctly divided into two regions: (1) Repetitive region that consists of an alternating alanine-rich and glycine-rich domain in which many prolines are present; and (2) C-terminal non-repetitive region. The region coding for 272 amino acids of MaSp gene was subcloned into prokaryotic expression vector pET28b(+) and an about 26kD recombinant protein was expressed at high levels in Escherichia coli BL21 (DE3) after induction of IPTG. After being purified with metal-affinity chromatography on Ni(2+) -IDA-Sepharose columns as well as gel filtration chromatography, the recombinant protein was confirmed to be predicted MaSp by means of amino acid composition analysis and N-terminal amino acid sequence analysis. The solubility behavior of recombinant MaSp with C-terminal non-repetitive region in the present study is similar to that of recombinant dragline silk proteins without C-terminal non-repetitive region expressed by bacteria and yeast in the other studies. The result shows that absence or presence of C-terminal non-repetitive region is not a crucial factor affecting the solubility of the recombinant MaSp.

To investigate the influence of the fermentation conditions on glycosylation of heterologous recombinant protein in yeast Pichia pastoris, the glycosylation of recombinant human interferon omega (rhIFNomega) under various fermentation conditions, e. g., cell density, initial pH, methanol concentration, duration of the induction, and medium volume were studied. The glycosylation of rhIFNomega in the continuous fermentation process under various pH values and in batch fermentation were also investigated. In 250 mL flask, the optimal cell density, initial pH, medium volume, methanol concentration and frequency of methanol induction were 250 g/L (WCW), pH6.0, less than 30 mL, 15 g/L and 3 (in every 24 h), respectively. In the continuous process, the glycosylation of rhIFNomega could be effectively improved by maintaining the pH value at 7.0-7.5. In the batch fermentation process, the expression level of glycosylated and non-glycosylated rhIFNomega were the same, but the specified value of glycosylation/non-glycosylation was significantly lower than that in the flask culture. The reason of this phenomenon will be further studied. This research lay the foundation for the scale-up of production and the enhancement of rhIFNomega glycosylation in Pichia pastoris.

The complete mitochondrial genome of Ophisaurus harti is a circular molecule of 17,163 bp in length, containing 13 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNAs and a control region. The A + T content of the overall base composition of H-strand is 54.9% (T: 23.4%; C: 30.8%; A: 31.5%; G: 14.3%). COI gene begins with GTG as start codon, while other 12 protein-coding genes start with a typical ATG initiation codon. ND1. COI. ATP8. ATP6. ND4L and ND5 genes are terminated with TAA as stop codon, ND2. ND6 and Cyt b end with TAG, COIII ends with TA, COII and ND3 end with T and ND4 ends with TG.

The complete mitochondrial genome of sika deer (Cervus nippon) South Anhui population is a circular molecule of 16,429 bp in length, containing 13 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNAs and a control region. The A + T content of the overall base composition of H-strand is 62% (T: 28.7%; C: 24.6%; A: 33.3%; G: 13.4%). ND2, ND3 and ND5 genes begin with ATA as start codon, ND4L gene begins with GTG as start codon, while other nine protein-coding genes start with a typical ATG initiation codon. ND1, COI, COII, ATP8, ATP6, ND4L, ND5 and ND6 genes are terminated with TAA as stop codon, ND2 ends with TAG, COIII, ND3 and ND4 ends with T, Cyt b ends with AGA.