Molecular Cloning and Characterization of a New C-type Lysozyme Gene from Yak Mammary Tissue

Jiang, Ming Feng,Hu, Ming Jun,Ren, Hong Hui,Wang, Li Asian Australasian Association of Animal Productio 2015 Animal Bioscience Vol.28 No.12

Milk lysozyme is the ubiquitous enzyme in milk of mammals. In this study, the cDNA sequence of a new chicken-type (c-type) milk lysozyme gene (YML), was cloned from yak mammary gland tissue. A 444 bp open reading frames, which encodes 148 amino acids (16.54 kDa) with a signal peptide of 18 amino acids, was sequenced. Further analysis indicated that the nucleic acid and amino acid sequences identities between yak and cow milk lysozyme were 89.04% and 80.41%, respectively. Recombinant yak milk lysozyme (rYML) was produced by Escherichia coli BL21 and Pichia pastoris X33. The highest lysozyme activity was detected for heterologous protein rYML5 (M = 1,864.24 U/mg, SD = 25.75) which was expressed in P. pastoris with expression vector $pPICZ{\alpha}A$ and it clearly inhibited growth of Staphylococcus aureus. Result of the YML gene expression using quantitative polymerase chain reaction showed that the YML gene was up-regulated to maximum at 30 day postpartum, that is, comparatively high YML can be found in initial milk production. The phylogenetic tree indicated that the amino acid sequence was similar to cow kidney lysozyme, which implied that the YML may have diverged from a different ancestor gene such as cow mammary glands. In our study, we suggest that YML be a new c-type lysozyme expressed in yak mammary glands that plays a role as host immunity.

Molecular Cloning and Expression of cDNA Encoding the Cysteine Proteinase Inhibitor from Upland Cotton

Ming-feng Jiang,Sheng-wei Li,Min Chen,Ying-fan Cai,Yong-fang Xie,Biao Li,Quan Sun,Huai-zhong Jiang,Zheng Pan,Yun-ling Gao,You-Lu Yuan,Yu-zheng Shi 한국식물학회 2009 Journal of Plant Biology Vol.52 No.5

A cDNA encoding a novel cysteine proteinase inhibitor (CPI) was isolated from a gland mutant Xiangmian-18 of upland cotton during the pigments gland forming stage. The cDNA comprises 378 bp and encodes 125 amino acid residues with molecular mass of 13.8 kDa. It contains the conserved motif of cysteine protease inhibitors and belongs to the cystatin superfamily (Gln- Val-Val-Ala-Gly). The deduced amino acid sequences of the domains are highly similar to the normal upland cotton (96.8%). SDS-PAGE and western hybridization analysis showed that the expressed recombinant protein was recombinant CPI. The inhibitory activity of recombinant CPI was 46 u/μg which was measured by inhibiting the protease activity of papain. RT-PCR results indicated that the expression level of developing gland stage was higher than that of undeveloped gland stage.

Modification of a Smoking Motivation Questionnaire for Chinese Medical Students

Jiang, Chao,Sun, Wen-Jie,Wan, Yan-Chun,Wei, Ming-Wei,Mu, Yong-Ping,Tarver, Siobhan L.,Gao, Yong-Qing,Hu, Tian,Xu, Chao,Gordon, James,Feng, Cindy Xin,Wen, Yu-Feng Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.6

Introduction: Smoking prevalence among the medical students is high in China. Therefore, understanding the smoking motivations of medical students is crucial for smoking control, but currently there are no scales questionnaires customized for probing the smoking motivations of medical students. This aim of study was to test and modify a questionnaire for investigating smoking motivations among medical students. Methods: A cross-sectional survey was conducted among 1,125 medical students at Xuzhou Medical College in China in 2012.The model fit and validity was assessed by confirmatory factor analysis (CFA) and the reliability was tested by single-item reliability, composite reliability, and item-total correlation. Results: The prevalence of smoking was 9.84 % among study population. In the modified scales, the global fit indices identified a CFI value of 0.96, TLI was 0.96, and the RMSEA was 0.063. CFA supported the two dimensional structure of the instrument. The average variance extracted ranged from 0.45 to 0.62. All single-item reliability scores were greater than 0.20, and the composite reliability ranged from 0.74 to 0.91. Conclusion: Modified scales could be the preliminary instrument used in evaluating the smoking motivations of medical students. However, it should be further assessed using other forms and methods of validity and reliability, additional motivations of smoking, and the survey of other medical colleges in China.

Cloning and characterization of a novel gene with alternative splicing in murine mesenchymal stem cell Line C3H/10T1/2 by gene trap screening

( Ming Ke Wang ),( Hui Qin Sun ),( Fan Jiang ),( Jing Han ),( Feng Ye ),( Tao Wang ),( Yong Ping Su ),( Zhong Min Zou ) 생화학분자생물학회 (구 한국생화학분자생물학회) 2010 BMB Reports Vol.43 No.12

A novel gene, designated mgt-6, containing four splicing variants, was isolated from a gene trap clone library of C3H/10T1/ 2 cells transfected with retroviral promoterless gene-trap vector, ROSAFARY. The transcript variants were differentially expressed in murine tissues and cell lines and differentially responded to diverse stimuli including TGF-β1 and mitogen-activated protein kinase (MAPK) inhibitors. The mgt-6 gene encoded a protein of 37 or 11 amino acid residuals with cytoplasmic distribution. However, when C3H/10T1/2 cells were treated with 5-azacytidine, the protein translocated into cell nucleus as indicated by fused LacZ or C-terminally tagged EGFP. Our preliminary results suggest that further study on the role of mgt-6 gene in cell transformation and differentiation may be of significance. [BMB reports 2010; 43(12): 789-794]

A multi-scale model linking microstructure and macro electric behaviors of ferroelectric field effect transistor

Limei Jiang,Xin Feng,Hao Ming,Qiong Yang,Jie Jiang,Min Liao 한국물리학회 2020 Current Applied Physics Vol.20 No.3

A multi-scale model linking microstructure and macro electric properties of ferroelectric field effect transistor (FeFET) is established based on the phase field method and Maxwell's total current law. The simulation results confirm that the channel current of FeFET is modulated by local interface ferroelectric domains which are easily affected by the microstructure. Using the developed transistor model, we investigate the effects of location of dislocation sites on hysteresis behavior of ferroelectric film and electric behavior of FeFET. Interfacial dislocations have strong effects on hysteresis behavior than internal dislocations since the former can induce imprint behavior while the latter only reduce the coercive field. However, due to the c-domains at the interface, the effects of interfacial and internal dislocations on FeFET electrical properties are the exact opposite. The interfacial dislocations just increase the voltage required to toggle the channel while the internal dislocations can induce disappearance of the FeFET memory window.

Off-Road Machinery Systms Engineering : Development of a Prototype of Guidance System for Rice-transplanter

( Fang Ming Zhang ),( Beom Soo Shin ),( Xi Ming Feng ),( Yuan Li ),( Ri Jiang Shou ) 한국농업기계학회 2013 바이오시스템공학 Vol.38 No.4

Purpose: It is not easy to drive a rice-transplanter avoiding underlapped or overlapped transplanting in paddy fields. An automated guidance system for the riding-type rice-transplanter would be necessary to operate the rice-transplanter autonomously or to assist the beginning drivers as a driving aid. Methods: A prototype of guidance system was composed of embedded computers, RTK-GPS, and a power-steering mechanism. Two Kalman filters were adopted to overcome sparse positioning data (1 Hz) from the RTK-GPS. A global Kalman filter estimated the posture of rice-transplanter every one second, and a local Kalman filter calculated the posture from every new estimation of the global Kalman filter with an interval of 200 ms. A PID controller was applied to the row-following mode control. A control method of U-turning mode was developed as well. A stepping motor with a reduction gear set was used to rotate the shaft of steering wheel. Results:Test trials for U-turning and row-following modes were done in a paddy field after some parameters have been tuned at the ground speed range of 0.3 ~ 1.2 m/s. The minimum RMS error of offset was 3.13 cm at the ground speed of 0.3 m/s while the maximum RMS error was 13.01 cm at 1.2 m/s. The offset RMS error tended to increase as the ground speed increased. The target point distance, LT also affected the system performance and PID controller parameters should be adjusted on different ground speeds. Conclusions: A target angle-based PID controller plus stationary steering angle controller made it possible for the rice-transplanter to steer autonomously by following a reference line accurately and even on U-turning mode. However, as condition in paddy fields is very complicated, the system should control the ground speed that prevents it from deviating too much due to ditch and slope.

Development of an automated guidance system for rice transplanter1

( Fang-ming Zhang ),( Xi-ming Feng ),( Yuan Li ),( Beom-soo Shin ),( Ru-jiang Shou ) 한국농업기계학회 2012 한국농업기계학회 학술발표논문집 Vol.17 No.1

An automated guidance system was developed on a rice transplanter. A distributed embedded computer system, one ECC and 5 ECUs, was constructed. Two Kalman filters were formed. The 1^st one, named as global Kalman filter, integrated data of a RTK-GPS, a strapdown attitude sensor, and two encoders. The 2^nd one, named as local Kalman filter, integrated data of a strapdown attitude sensor and two encoders. A PID controller was applied to row following control, whose offset RMS could less than 4 cm at speed of 1.2 m/s. A U-turn controller was also designed that it could control the transplanter in a go-turn-back mode.

Development of a Prototype of Guidance System for Rice-transplanter

Zhang, Fang-Ming,Shin, Beom-Soo,Feng, Xi-Ming,Li, Yuan,Shou, Ru-Jiang Korean Society for Agricultural Machinery 2013 바이오시스템공학 Vol.38 No.4

Purpose: It is not easy to drive a rice-transplanter avoiding underlapped or overlapped transplanting in paddy fields. An automated guidance system for the riding-type rice-transplanter would be necessary to operate the rice-transplanter autonomously or to assist the beginning drivers as a driving aid. Methods: A prototype of guidance system was composed of embedded computers, RTK-GPS, and a power-steering mechanism. Two Kalman filters were adopted to overcome sparse positioning data (1 Hz) from the RTK-GPS. A global Kalman filter estimated the posture of rice-transplanter every one second, and a local Kalman filter calculated the posture from every new estimation of the global Kalman filter with an interval of 200 ms. A PID controller was applied to the row-following mode control. A control method of U-turning mode was developed as well. A stepping motor with a reduction gear set was used to rotate the shaft of steering wheel. Results: Test trials for U-turning and row-following modes were done in a paddy field after some parameters have been tuned at the ground speed range of 0.3 ~ 1.2 m/s. The minimum RMS error of offset was 3.13 cm at the ground speed of 0.3 m/s while the maximum RMS error was 13.01 cm at 1.2 m/s. The offset RMS error tended to increase as the ground speed increased. The target point distance, LT also affected the system performance and PID controller parameters should be adjusted on different ground speeds. Conclusions: A target angle-based PID controller plus stationary steering angle controller made it possible for the rice-transplanter to steer autonomously by following a reference line accurately and even on U-turning mode. However, as condition in paddy fields is very complicated, the system should control the ground speed that prevents it from deviating too much due to ditch and slope.

Cloning and Expression of Yak Active Chymosin in Pichia pastoris

Luo, Fan,Jiang, Wei Hua,Yang, Yuan Xiao,Li, Jiang,Jiang, Ming Feng Asian Australasian Association of Animal Productio 2016 Animal Bioscience Vol.29 No.9

Rennet, a complex of enzymes found in the stomachs of ruminants, is an important component for cheese production. In our study, we described that yak chymosin gene recombinant Pichia pastoris strain could serve as a novel source for rennet production. Yaks total RNA was extracted from the abomasum of an unweaned yak. The yak preprochymosin, prochymosin, and chymosin genes from total RNA were isolated using gene specific primers based on cattle chymosin gene sequence respectively and analyzed their expression pattern byreal time-polymerase chain reaction. The result showed that the chymosin gene expression level of the sucking yaks was 11.45 times higher than one of adult yaks and yak chymosin belongs to Bovidae family in phylogenetic analysis. To express each, the preprochymosin, prochymosin, and chymosin genes were ligated into the expression vector $pPICZ{\alpha}A$, respectively, and were expressed in Pichia pastoris X33. The results showed that all the recombinant clones of P. pastoris containing the preprochymosin, prochymosin or chymosin genes could produce the active form of recombinant chymosin into the culture supernatant. Heterologous expressed prochymosin (14.55 Soxhlet unit/mL) had the highest enzyme activity of the three expressed chymosin enzymes. Therefore, we suggest that the yak chymosin gene recombinant Pichia pastoris strain could provide an alternative source of rennet production.

노인복지관의 치유환경 디자인 특성에 관한 연구

곽풍명(Guo, Feng-Ming),강조양(Jiang, Zhao-Yang),상금(Shang, Jin),황연숙(Hwang, Yeon-Sook) 한국실내디자인학회 2022 한국실내디자인학회 학술대회논문집 Vol.24 No.1