Co-expression of recombinant human prolyl with human collagen α1 (III) chains in two yeast systems.

UNLABELLED: In this study, we co-expressed the human prolyl 4-hydroxylases (P4H) with human collagen α1 (III) (COL3A1) in an inducible system: Pichia pastoris (pPICZB), and one constitutive system: P. pastoris (pGAPZαB). The P4H catalyses the post-translational hydroxylation of proline residues in collagen strands. Conventional protein expression system such as bacteria and yeasts, which lack endogenous P4H, are not efficient for the production of recombinant collagen. In this study, the P4H gene was constructed in pGAPZαB plasmid and pPICZB plasmid respectively. These two plasmids were transformed in P. pastoris #1 that carrying COL3A1. Colony PCR analysis and sequencing after electroporation P. pastoris GS115 showed that the target gene had inserted successfully. The results of reverse transcript-qPCR, SDS-PAGE, Western blotting and LC-MS/MS analysis of the rhCOL3A1 demonstrated that the P4H was expressed successfully. Besides, it is noted that low copy number, constitutive system was suitable for hydroxylated rhCOL3A1. SIGNIFICANCE AND IMPACT OF THE STUDY: Successful co-expression of recombinant human collagen α1 (III) (rhCOL3A1) and human prolyl 4-hydroxylases (P4H) in Picha pastoris GS115, simultaneously results in the acquisition of rhCOL3A1 with hydroxylation of proline (Hyp). Further, this experiment also discusses that the high or low copy numbers and different promoters affect the Hyp degree of rhCOL3A1. Selecting more appropriate strains can express high degree Hyp of rhCOL3A1. This work will be helpful to the collagen structure study.

Survey of sweetpotato viruses in China.

Ten sweetpotato viruses were surveyed in 3 major sweetpotato planting region (covering 11 provinces) in China from 2006 to 2010 to understand the distribution of sweetpotato viral diseases. Nine out of the 10 viruses were found in every major planting region. The most frequently detected virus in the Northern and the Yangtze River region was SPMSV. In the Southern region, SPVG was the most frequently detected virus. Compared to the results of the survey done in 1989, the incidences of all the viral diseases increased.

[Role of the ES-62 protein derived from Acanthocheilonema viteae in regulation of immune dysregulation diseases: a review].

ES-62 is a phosphorylcholine-containing, 62 kDa glycoprotein derived from the excretory-secretory product of Acanthocheilonema viteae, which is effective for the prevention and treatment of immune dysregulation diseases through triggering activation of immune cells, such as dendritic cells, mononuclear macrophages and regulatory B cells and mediating immune responses. Recently, the role of the ES-62 protein in the management of allergic, autoimmune and metabolic diseases has been paid much attention. This review summarizes the regulatory role of the ES-62 protein in immune dysregulation diseases and the underlying mechanisms, so as to provide insights into future experimental studies.

Peanut (Arachis hypogaea L.) S-adenosylmethionine decarboxylase confers transgenic tobacco with elevated tolerance to salt stress.

Polyamines play an important role in stress response. In the pathway of polyamines synthesis, S-adenosylmethionine decarboxylase (SAMDC) is one of the key enzymes. In this study, a full length cDNA of SAMDC (AhSAMDC) was isolated from peanut (Arachis hypogaea L.). Phylogenetic analysis revealed high sequence similarity between AhSAMDC and SAMDC from other plants. In peanut seedlings exposed to sodium chloride (NaCl), the transcript level of AhSAMDC in roots was the highest at 24 h that decreased sharply at 72 and 96 h after 150 mM NaCl treatment. However, the expression of AhSAMDC in peanut leaves was significantly inhibited, and the transcript levels in leaves were not different compared with control These results implied the tissue-specific and time-specific expression of AhSAMDC. The physiological effects and functional mechanism of AhSAMDC were further evaluated by overexpressing AhSAMDC in tobaccos. The transgenic tobacco lines exhibited higher germination rate and longer root length under salt stress. Reduced membrane damage, higher antioxidant enzyme activity, and higher proline content were also observed in the transgenic tobacco seedlings. What's more, AhSAMDC also led to higher contents of spermidine and spermine, which can help to scavenge reactive oxygen species. Together, this study suggests that AhSAMDC enhances plant resistance to salt stress by improving polyamine content and alleviating membrane damage.

[Analysis of components of proteins from Echinococcus granulosus cyst fluid].

OBJECTIVE: To analyze the components of proteins from Echinococcus granulosus cyst fluid using the shotgun method, and to identify the active components with potential regulatory effects for immune dysregulation diseases. METHODS: The E. granulosus cyst fluid was collected aseptically from the hepatic cysts of patients with cystic echinococcosis, and characterized by liquid chromatography (LC) tandem mass spectrometry (MS/MS) following digestion with trypsin. The protein data were searched using the software MaxQuant version 1.6.1.0 and the cellular components, molecular functions, and biological processes of the identified proteins were analyzed using the Gene Ontology (GO) method. RESULTS: The E. granulosus cyst fluid separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) had a relative molecular mass of 25 to 70 kDa. LS-MS/MS analysis identified 37 proteins, including 32 known proteins and 5 unknown proteins. At least 4 proteins were preliminarily found to exhibit potential regulatory effects for immune dysregulation diseases, including antigen B, glutathione-S-transferase (GST), thioredoxin peroxidase (TPX) and malate dehydrogenase (MDH). GO enrichment analysis showed that the identified proteins had 149 molecular functions and were involved in 341 biological processes. CONCLUSIONS: E. granulosus cyst fluid has a variety of protein components, and four known proteins are preliminarily identified to be associated with immune dysregulation diseases.