By PCR method, apo phycoerythrocyanin α subunit gene (pecA) of Mastigocladus laminosus (M. laminosus) was amplified from its genomic DNA, and then cloned in pBluescript. The pecA gene was subcloned into the exp...By PCR method, apo phycoerythrocyanin α subunit gene (pecA) of Mastigocladus laminosus (M. laminosus) was amplified from its genomic DNA, and then cloned in pBluescript. The pecA gene was subcloned into the expression vector pGEMD, and then transformed into E.coli BL21 (DE3). After induction, a new protein of molecular weight 19×10 3 existing in inclusion body was overexpressed. The expressed product was confirmed to be apo phycoerythrocyanin α subunit by Dot ELISA.展开更多
This study investigated the abnormal expression of ATP synthase β-subunit（ATPsyn-β） in pancreas islets of rat model of polycystic ovary syndrome（PCOS） with type 2 diabetes mellitus（T2DM）,and the secretion func...This study investigated the abnormal expression of ATP synthase β-subunit（ATPsyn-β） in pancreas islets of rat model of polycystic ovary syndrome（PCOS） with type 2 diabetes mellitus（T2DM）,and the secretion function changes after up-regulation of ATP5 b.Sixty female SD rats were divided into three groups randomly and equally.The rat model of PCOS with T2 DM was established by free access to the high-carbohydrate/high-fat diet,subcutaneous injections of DHEA,and a single injection of streptozotocin.The pancreas was removed for the detection of the ATPsyn-β expression by immunohistochemical staining,Western blotting and reverse transcription-PCR（RT-PCR）.The pancreas islets of the rats were cultured,isolated with collagenase Ⅴ and purified by gradient centrifugation,and the insulin secretion after treatment with different glucose concentrations was tested.Lentivirus ATP5 b was successfully constructed with the vector of GV208 and transfected into the pancreas islets for the over-expression of ATPsyn-β.The insulin secretion and intracellular ATP content were determined after transfection of the PCOS-T2 DM pancreas islets with Lenti-ATP5 b.The results showed that the expression of ATPsyn-β protein and m RNA was significantly decreased in the pancreas of PCOS-T2 DM rats.The ATP content in the pancreas islets was greatly increased and the insulin secretion was improved after the up-regulation of ATPsyn-β in the pancreas islets transfected with lenti-ATP5 b.These results indicated that for PCOS,the ATPsyn-β might be one of the key factors for the attack of T2 DM.展开更多
The effects of the deficiency of the allergenicα-subunit on soybean amino acid(AA)composition were studied using the cultivar Dongnong 47(DN47)as a genetic background.The near-isogenic line(NIL)NIL-DN47-Δα of DN47,...The effects of the deficiency of the allergenicα-subunit on soybean amino acid(AA)composition were studied using the cultivar Dongnong 47(DN47)as a genetic background.The near-isogenic line(NIL)NIL-DN47-Δα of DN47,with an introgression of theα-null trait allele from the high protein donor parent RiB,was created by marker assisted background selection and used to investigate the AA content and nutritional quality.The contents of crude protein,the total AAs,the total essential amino acids(EAAs)and sulfur-containing(Met and Cys)AAs increased by 4.11%,4.16%,5.20% and 11.96%,respectively in NIL-DN47-Δα compared with DN47.Analyses of the total EAAs(TEAAs)and the EAA index(EAAI)revealed that both parameters in NIL-DN47-Δα were higher than those in DN47.The null-allele of theα-subunit positively affected the AA scores.The quantitative changes in free AAs(FAAs)in the developing seeds of NIL-DN47-Δα and DN47 were compared as of 15 days after flowering(DAF)until maturity.The results showed that the total FAA content in NIL-DN47-Δα was significantly higher than that in the DN47 throughout the late maturation stage(40-60 DAF)of seeds.The high concentration of the FAAs in cgy-2 mutant seeds was a consequence of the high rates of synthesis and/or accumulation of individual FAAs during seed maturation where 25 DAF was an important turning point in the accumulation of the FAAs.The FAA contents of single soybeanα-null,double(α+α′)and triple(α+α′+group I)-null mutant combination lines were investigated.In all of these combinations,introduction of the cgy-2 gene invariably raised the FAA content of mature seeds above that of the DN47.In summary,the enhanced protein quality in cgy-2 mutants resulted from several factors.(1)There was a general increase in the contents of most AAs and FAAs in NIL-DN47-Δα.(2)The induced synthesis of free Arg contributed effectively to the high FAAs of various storage-protein-deficiency mutants.For example,in the S2(null α,group I),the free Arg content was seven times as much as that of DN47,accounting for more than half of the total FAA content in the seed.(3)The increase of sulfur-containing AAs in theα-null type NIL mainly resulted from elevated Met content.These data suggested that the cgy-2 mutation might improve the protein quality of soybean seeds and that lacked of the allergenicα-subunit resulted in increased the FAA content.展开更多
In this study, the immuno-response of recombinant hCG-β and natural hCGβ was comparatively investigated by using Freund's adjuvant. The results showed that, the properties and merits of the antibodies elicited b...In this study, the immuno-response of recombinant hCG-β and natural hCGβ was comparatively investigated by using Freund's adjuvant. The results showed that, the properties and merits of the antibodies elicited by both kinds of hCG-β were similar. The antisera had high affinity for binding with hCG (Kαγβ≈5.86×108/mol/L, Kαβ≈8.18×108/mol/L), and were found to be effective in inhibiting the binding of 125I-hCG to receptors in rat testes. Results also indicated that, similar to the antisera induced by natural hCG-β, the recombinant hCG-β induced antisera had capacity of neutralizing the biological activities of hCG. Recombinant hCG-β could be used as an immunogen for contraceptive vaccine.展开更多
CACNA1 S gene is the gene encoding L-type calcium channel αa-subunit. CACNA1 S gene mutations can cause hypokalemic periodic pa- ralysis （HOKPP）. The related research speculated that CACNA1 S gene was the candidate...CACNA1 S gene is the gene encoding L-type calcium channel αa-subunit. CACNA1 S gene mutations can cause hypokalemic periodic pa- ralysis （HOKPP）. The related research speculated that CACNA1 S gene was the candidate genes which affect meat quality traits. In the present ar- ticle, the biological characteristics of CACNA1 S gene, structure, genetic diseases and the research development were respectively reviewed so as to provide a reference for further research.展开更多
文摘By PCR method, apo phycoerythrocyanin α subunit gene (pecA) of Mastigocladus laminosus (M. laminosus) was amplified from its genomic DNA, and then cloned in pBluescript. The pecA gene was subcloned into the expression vector pGEMD, and then transformed into E.coli BL21 (DE3). After induction, a new protein of molecular weight 19×10 3 existing in inclusion body was overexpressed. The expressed product was confirmed to be apo phycoerythrocyanin α subunit by Dot ELISA.
文摘This study investigated the abnormal expression of ATP synthase β-subunit（ATPsyn-β） in pancreas islets of rat model of polycystic ovary syndrome（PCOS） with type 2 diabetes mellitus（T2DM）,and the secretion function changes after up-regulation of ATP5 b.Sixty female SD rats were divided into three groups randomly and equally.The rat model of PCOS with T2 DM was established by free access to the high-carbohydrate/high-fat diet,subcutaneous injections of DHEA,and a single injection of streptozotocin.The pancreas was removed for the detection of the ATPsyn-β expression by immunohistochemical staining,Western blotting and reverse transcription-PCR（RT-PCR）.The pancreas islets of the rats were cultured,isolated with collagenase Ⅴ and purified by gradient centrifugation,and the insulin secretion after treatment with different glucose concentrations was tested.Lentivirus ATP5 b was successfully constructed with the vector of GV208 and transfected into the pancreas islets for the over-expression of ATPsyn-β.The insulin secretion and intracellular ATP content were determined after transfection of the PCOS-T2 DM pancreas islets with Lenti-ATP5 b.The results showed that the expression of ATPsyn-β protein and m RNA was significantly decreased in the pancreas of PCOS-T2 DM rats.The ATP content in the pancreas islets was greatly increased and the insulin secretion was improved after the up-regulation of ATPsyn-β in the pancreas islets transfected with lenti-ATP5 b.These results indicated that for PCOS,the ATPsyn-β might be one of the key factors for the attack of T2 DM.
基金Support by the National Natural Science Foundation of China(31371650,31071440)Northeast Agricultural University Innovation Foundation For Postgraduates(yjscx4042)。
文摘The effects of the deficiency of the allergenicα-subunit on soybean amino acid(AA)composition were studied using the cultivar Dongnong 47(DN47)as a genetic background.The near-isogenic line(NIL)NIL-DN47-Δα of DN47,with an introgression of theα-null trait allele from the high protein donor parent RiB,was created by marker assisted background selection and used to investigate the AA content and nutritional quality.The contents of crude protein,the total AAs,the total essential amino acids(EAAs)and sulfur-containing(Met and Cys)AAs increased by 4.11%,4.16%,5.20% and 11.96%,respectively in NIL-DN47-Δα compared with DN47.Analyses of the total EAAs(TEAAs)and the EAA index(EAAI)revealed that both parameters in NIL-DN47-Δα were higher than those in DN47.The null-allele of theα-subunit positively affected the AA scores.The quantitative changes in free AAs(FAAs)in the developing seeds of NIL-DN47-Δα and DN47 were compared as of 15 days after flowering(DAF)until maturity.The results showed that the total FAA content in NIL-DN47-Δα was significantly higher than that in the DN47 throughout the late maturation stage(40-60 DAF)of seeds.The high concentration of the FAAs in cgy-2 mutant seeds was a consequence of the high rates of synthesis and/or accumulation of individual FAAs during seed maturation where 25 DAF was an important turning point in the accumulation of the FAAs.The FAA contents of single soybeanα-null,double(α+α′)and triple(α+α′+group I)-null mutant combination lines were investigated.In all of these combinations,introduction of the cgy-2 gene invariably raised the FAA content of mature seeds above that of the DN47.In summary,the enhanced protein quality in cgy-2 mutants resulted from several factors.(1)There was a general increase in the contents of most AAs and FAAs in NIL-DN47-Δα.(2)The induced synthesis of free Arg contributed effectively to the high FAAs of various storage-protein-deficiency mutants.For example,in the S2(null α,group I),the free Arg content was seven times as much as that of DN47,accounting for more than half of the total FAA content in the seed.(3)The increase of sulfur-containing AAs in theα-null type NIL mainly resulted from elevated Met content.These data suggested that the cgy-2 mutation might improve the protein quality of soybean seeds and that lacked of the allergenicα-subunit resulted in increased the FAA content.
文摘In this study, the immuno-response of recombinant hCG-β and natural hCGβ was comparatively investigated by using Freund's adjuvant. The results showed that, the properties and merits of the antibodies elicited by both kinds of hCG-β were similar. The antisera had high affinity for binding with hCG (Kαγβ≈5.86×108/mol/L, Kαβ≈8.18×108/mol/L), and were found to be effective in inhibiting the binding of 125I-hCG to receptors in rat testes. Results also indicated that, similar to the antisera induced by natural hCG-β, the recombinant hCG-β induced antisera had capacity of neutralizing the biological activities of hCG. Recombinant hCG-β could be used as an immunogen for contraceptive vaccine.
文摘CACNA1 S gene is the gene encoding L-type calcium channel αa-subunit. CACNA1 S gene mutations can cause hypokalemic periodic pa- ralysis （HOKPP）. The related research speculated that CACNA1 S gene was the candidate genes which affect meat quality traits. In the present ar- ticle, the biological characteristics of CACNA1 S gene, structure, genetic diseases and the research development were respectively reviewed so as to provide a reference for further research.
