Objective： The aim of the study was to observe the transfection efficacy of hepatitis B virus envelope （HBVE） and evaluate its ability as a gene transfer vector for liver cancer cells. Methods： To obtain HBVE, the...Objective： The aim of the study was to observe the transfection efficacy of hepatitis B virus envelope （HBVE） and evaluate its ability as a gene transfer vector for liver cancer cells. Methods： To obtain HBVE, the supematant fluid of HepG 2.2.15 cells was mixed with a PEG8000 solution for concentration and was inactivated by β-propiolactone. The acquired HBVE was used to pack plRES2-EGFP to test its package ability. Then, we examined its quantity and quality with ELISA, PCR, SDS-PAGE and electron microscopy. The plRES2-EGFP was packed with HBVE and obtained the product HBVE-GFP. The plRES2-EGFP was packed with liposome and obtained the product liposome-GFP. HBVE-GFP and liposome-GFP were used to transfer HepG 2 cells to study the transfection efficiency. HBVE-GFP was used to transfer HepG 2, A549, HeLa and FB cells to study the targeting ability. The green fluorescent protein （GFP） expression was observed under a fluorescent microscope. The rate of GFP positive cells was determined by flow cytometry. Results： 1. The acquired HBVE could retain the surface protein HBsAg ＋ pre S1 ＋ pre S2 and had no virus DNA. It had good package ability for plRES2-EGFP. 2. Transfection efficiency： The GFP could be observed in both the liposome group and HBVE group under the fluorescent microscope. But the HBVE group had a higher fluorescent intensity than liposome group. The transfection rate of liposome group was 49.97% ＋ 2.37% while the HBVE group was 70.65% ＋ 3.15% and the fluorescent intensity of the HBVE group was 3-4 times （P = 0.000） for liposome group with the determination of flow cytometry. 3. Targeting ability： The GFP could be observed in the four groups under the fluorescent microscope. The HepG 2 group had the highest fluorescent intensity among the four groups. The transfection rate of HepG 2 group was 71.35% ＋ 0.03% which was highly expressed than other groups （P = 0.000） and the fluorescent intensity of the HepG 2 group was 2-3 times （P = 0.000） for the other 3 groups with the determination of flow cytometry. Conclusion： HBVE can be constructed successfully with the methods of PEG8000 and β-propiolactone from the supernatant fluid of HepG 22.15 cells. The HBVE can be a candidate gene transfer vector for liver cancer cells.展开更多
[Objective] This research aimed at constructing two gateway-compatible plant expression vectors for functional genomics of abiotic stress in tomato.[Method] pK2GW7I was generated from the plant expression vector pK2GW...[Objective] This research aimed at constructing two gateway-compatible plant expression vectors for functional genomics of abiotic stress in tomato.[Method] pK2GW7I was generated from the plant expression vector pK2GW7,0 by replacing the CaMV 35S promoter with abiotic-stress inducible plant promoter pRD29A,which was derived from the promoter of Arabidopsis gene RD29A.pKGW121 was generated by replacing the CaMV 35S of the plant expression vector pRD410 with the gateway recombinant cassette(attR1-cmR-ccdB-attR2)from pK2GW7,0.Constitutive and root-specific promoters were tested on pKGW121.[Result] Two gateway-compatible destination vectors,pK2GW7I and pKGW121,were successfully constructed;practicability test proved that pKGW121 was applicable for promoter analysis.[Conclusion] With the incorporation of gateway cloning technology or abiotic-stress inducible promoter,pKGW121 and pK2GW7I will be promising in large-scale investigation of tomato functional genes associated with various abiotic stresses.A high-throughput workflow for the construction of plant transformation vector was proposed and validated.展开更多
To solve the problem of stray interference to star point target identification while a star sensor imaging to the sky, a study on space luminous environment adaptability of missile-borne star sensor was carried out. B...To solve the problem of stray interference to star point target identification while a star sensor imaging to the sky, a study on space luminous environment adaptability of missile-borne star sensor was carried out. By Plank blackbody radiation law and some astronomic knowledge, irradiancies of the stray at the star sensor working height were estimated. By relative astrophysical and mathematics knowledge, included angles between the star sensor optical axis point and the stray at any moment were calculated. The calculation correctness was verified with the star map software of Stellarium. By combining the upper analysis with the baffle suppression effect, a real-time model for space luminous environment of missile-borne star sensor was proposed. By signal-noise rate （SNR） criterion, the adaptability of missile-borne star sensor to space luminous environment was studied. As an example, a certain type of star sensor was considered when imaging to the starry sky on June 22, 2011 （the Summer Solstice） and September 20, 2011 （August 23 of the lunar year, last quarter moon） in Beijing. The space luminous environment and the adaptability to it were simulated and analyzed at the star sensor working height. In each period of time, the stray suppression of the baffle is analyzed by comparing the calculated included angle between the star sensor optical axis point and the stray with the shielded provided by system index. When the included angle is larger than the shielded angle and less than 90~, the stray is restrained by the baffle. The stray effect on star point target identification is analyzed by comparing the irradiancy of 6 magnitude star with that of the stray on star sensor sensitization surface. When the irradiancy of 6 magnitude star is 5 times more than that of the stray, there is no effect on the star point target identification. The simulation results are identicat with the actual situation. The space luminous environment of the missile-borne star sensor can be estimated real-timely by this model. The adaptability of the star sensor to space luminous environment can be analyzed conveniently. A basis for determining the relative star sensor indexes, the navigation star chosen strategy and the missile launch window can be provided.展开更多
基金Supported by a grant from the National Natural Sciences Foundation of China No 30100189
文摘Objective： The aim of the study was to observe the transfection efficacy of hepatitis B virus envelope （HBVE） and evaluate its ability as a gene transfer vector for liver cancer cells. Methods： To obtain HBVE, the supematant fluid of HepG 2.2.15 cells was mixed with a PEG8000 solution for concentration and was inactivated by β-propiolactone. The acquired HBVE was used to pack plRES2-EGFP to test its package ability. Then, we examined its quantity and quality with ELISA, PCR, SDS-PAGE and electron microscopy. The plRES2-EGFP was packed with HBVE and obtained the product HBVE-GFP. The plRES2-EGFP was packed with liposome and obtained the product liposome-GFP. HBVE-GFP and liposome-GFP were used to transfer HepG 2 cells to study the transfection efficiency. HBVE-GFP was used to transfer HepG 2, A549, HeLa and FB cells to study the targeting ability. The green fluorescent protein （GFP） expression was observed under a fluorescent microscope. The rate of GFP positive cells was determined by flow cytometry. Results： 1. The acquired HBVE could retain the surface protein HBsAg ＋ pre S1 ＋ pre S2 and had no virus DNA. It had good package ability for plRES2-EGFP. 2. Transfection efficiency： The GFP could be observed in both the liposome group and HBVE group under the fluorescent microscope. But the HBVE group had a higher fluorescent intensity than liposome group. The transfection rate of liposome group was 49.97% ＋ 2.37% while the HBVE group was 70.65% ＋ 3.15% and the fluorescent intensity of the HBVE group was 3-4 times （P = 0.000） for liposome group with the determination of flow cytometry. 3. Targeting ability： The GFP could be observed in the four groups under the fluorescent microscope. The HepG 2 group had the highest fluorescent intensity among the four groups. The transfection rate of HepG 2 group was 71.35% ＋ 0.03% which was highly expressed than other groups （P = 0.000） and the fluorescent intensity of the HepG 2 group was 2-3 times （P = 0.000） for the other 3 groups with the determination of flow cytometry. Conclusion： HBVE can be constructed successfully with the methods of PEG8000 and β-propiolactone from the supernatant fluid of HepG 22.15 cells. The HBVE can be a candidate gene transfer vector for liver cancer cells.
基金Supported by the Natural Science Foundation of China(30771461)the Wuhan Chenguang Program of Hubei Provicne of China(20055003059-24)
文摘[Objective] This research aimed at constructing two gateway-compatible plant expression vectors for functional genomics of abiotic stress in tomato.[Method] pK2GW7I was generated from the plant expression vector pK2GW7,0 by replacing the CaMV 35S promoter with abiotic-stress inducible plant promoter pRD29A,which was derived from the promoter of Arabidopsis gene RD29A.pKGW121 was generated by replacing the CaMV 35S of the plant expression vector pRD410 with the gateway recombinant cassette(attR1-cmR-ccdB-attR2)from pK2GW7,0.Constitutive and root-specific promoters were tested on pKGW121.[Result] Two gateway-compatible destination vectors,pK2GW7I and pKGW121,were successfully constructed;practicability test proved that pKGW121 was applicable for promoter analysis.[Conclusion] With the incorporation of gateway cloning technology or abiotic-stress inducible promoter,pKGW121 and pK2GW7I will be promising in large-scale investigation of tomato functional genes associated with various abiotic stresses.A high-throughput workflow for the construction of plant transformation vector was proposed and validated.
文摘To solve the problem of stray interference to star point target identification while a star sensor imaging to the sky, a study on space luminous environment adaptability of missile-borne star sensor was carried out. By Plank blackbody radiation law and some astronomic knowledge, irradiancies of the stray at the star sensor working height were estimated. By relative astrophysical and mathematics knowledge, included angles between the star sensor optical axis point and the stray at any moment were calculated. The calculation correctness was verified with the star map software of Stellarium. By combining the upper analysis with the baffle suppression effect, a real-time model for space luminous environment of missile-borne star sensor was proposed. By signal-noise rate （SNR） criterion, the adaptability of missile-borne star sensor to space luminous environment was studied. As an example, a certain type of star sensor was considered when imaging to the starry sky on June 22, 2011 （the Summer Solstice） and September 20, 2011 （August 23 of the lunar year, last quarter moon） in Beijing. The space luminous environment and the adaptability to it were simulated and analyzed at the star sensor working height. In each period of time, the stray suppression of the baffle is analyzed by comparing the calculated included angle between the star sensor optical axis point and the stray with the shielded provided by system index. When the included angle is larger than the shielded angle and less than 90~, the stray is restrained by the baffle. The stray effect on star point target identification is analyzed by comparing the irradiancy of 6 magnitude star with that of the stray on star sensor sensitization surface. When the irradiancy of 6 magnitude star is 5 times more than that of the stray, there is no effect on the star point target identification. The simulation results are identicat with the actual situation. The space luminous environment of the missile-borne star sensor can be estimated real-timely by this model. The adaptability of the star sensor to space luminous environment can be analyzed conveniently. A basis for determining the relative star sensor indexes, the navigation star chosen strategy and the missile launch window can be provided.
