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The polar tube protein is the major component of polar tube, and can specifically locate on the polar tube of microsporidia and plays an important role in invasion host cell. In this study, we analyzed the potential O- and Nglycosylation sites in polar tube protein 1 from Nosema bombycis. NbPTP1 was successfully cloned to eukaryotic expression vector pMT/Bip/V5-His A, involved V5 and His tags. After transfection, NbPTP1 gene could be efficiently expressed in Drosophila S2 cells. In addition, Lectin blotting and beta elimination analysis showed that NbPTP1 expressed in Drosophila S2 cells was O-glycosylation. These studies provided a basis for understanding the relationship between glycosylation and function of NbPTP1, helped us to reveal the infection mechanism of microsporidia and established effective diagnosis and prevention methods for microsporidia.
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Objective To explore the correlation between axon guidance factor Semaphorin 5A and clinicopathological features and its role in the invasion and metastasis of gastric cancer.Methods The expression of Semaphorin 5A in gastric cancer tissues of 171 patients with different gender,age,histological type and TNM stage was detected with immunohistochemistry assay.The expression of Semaphorin 5A was determined by Western blotting assay in gastric cancer cell lines SGC7901 and MKN-45 with metastatic ability and gastric cancer cell lines SNU-1 and AGS without metastatic ability.With RNA interfere technique(RNAi),Semaphorin 5A siRNA expression vector was constructed and transfected into gastric cancer cell line SGC7901.The stable gastric cancer cell line down-expressing Semaphorin 5A was established.The effect of Semaphorin 5A gene silencing on the adhesion,migration and invasion of gastric cancer cell was examined by cell adhesion test,wound healing test and transwell chamber assays.Results The expression level of Semaphorin 5A was correlated with the differentiation degree of gastric cancer(x2 =6.32,P =0.01),lymphnode metastasis(x2 =7.68,P=0.01)and distant metastasis of gastric cancer(x2 =13.67,P =0.00),not correlated with age(x2 =0.21,P=0.79),gender(x2=1.79,P=0.15)and the depth of gastric cancer invasion(x2=1.34,P=0.55).The expression of Semaphorin 5A in cell lines SGC7901 and MKN-45 was significantly higher than that of cell lines SNU-1 and AGS(P＜0.01).Semaphorin 5A gene silencing significantly suppressed the adhesion,migration and invasion abilities of gastric cancer cells.Conclusion Semaphorin 5A may play a catalytic role in the invasion and metastasis of gastric cancer through increasing the adhesion,migration and invasion abilities of gastric cancer cell.
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Recently, more research about the plant bioreactor expressing genes encoding human proteins was reported. In the present study, the cDNA of the human gene keratinocyte growth factor 2 (KGF2) was replaced with plant preferred codons by PCR, and the modified full-length cDNA was cloned into the plant expression vector pCAMBIA-YO containing the oil-body promoter. The fusion construct pCAMBIA-YO-KGF2 was transformed into Brassica napus by Agrobacterium tumefacien-mediated cotyledon transformation method. The transgenic seedlings were identified by PCR, Southern and western blot analysis all showed that KGF2 gene was successfully expressed in in transgenic Brassica napus.
Subject(s)
Humans , Brassica napus , Genetics , Metabolism , Cloning, Molecular , DNA, Complementary , Genetics , Fibroblast Growth Factor 7 , Genetics , Genetic Vectors , Genetics , Plants, Genetically Modified , Genetics , Rhizobium , Genetics , Transformation, GeneticABSTRACT
0.05) ; whereas it was seen in CNE-2Z-H5-9. Anti-PAI-1 antibody did not inhibit the heterogeneous adhesion and invsion of CNE-2Z-H5-9. Conclusions:PAI-1 maybe inhibit the metastasis and invasion of NPC.
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Purpose:To study the expression of uPA, uPAR ,PAI 1 in 3 nasopharyngeal carcinoma cell lines CEN 2Z,CEN 2Z H5,CEN 2Z H5 9,with different ability to metastasize through lymph.Methods:RT PCR was used to detect expression of uPA,uPAR,PAI 1 at levels of mRNA.Results:mRNA expression of uPA,uPAR was the highest in CNE 2Z H5 9,whereas the lowest in CNE 2Z;mRNA expression of PAI 1was detected in CNE 2Z and CNE 2Z H5, but revealed no differences,but not in CNE 2Z H5 9.Conclusions:uPA?uPAR may promote the metastasis and invasion of NPC,whereas PAI 1 maybe inhibit it