基于适冷酶研究的生物综合实验教学设计
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摘要
通过引入专业热点问题,并结合学校海洋培养特色,生物综合实验教学设计以海洋微生物适冷酶为材料,对其基因的克隆与生物信息学进行分析。采用同源建模分析手段,分析适冷酶的结构特点;利用亲和层析方法对重组适冷酶进行分离纯化; SDS-PAGE分析基因工程产物及纯度鉴定,研究重组蛋白活性。通过综合实验使学生深入了解海洋微生物适冷酶基因,挖掘学生的创新潜力,提升学生的实验操作技能,进而达到提高实验教学质量的目的。
Through the introduction of scientific issues,and in combination with the marine training features of Harbin Institute of Technology, the genes of cold-adapted enzymes from marine microorganisms were cloned and the bioinformatics were analyzed in the comprehensive biology experiment. Then the structure characteristics of cold-adapted enzymes were also analyzed using homology modeling. The recombinant enzyme was purified using affinity chromatography. Besides,the genetic engineering products and identify the purity were analyzed by SDS-PAGE. The recombinant enzyme activities were also determined. The comprehensive biology experiment could make students a deep understanding of the research on the genes of cold-adapted enzymes from marine microorganisms,improved their potential for innovation and experimental operation skills,and then achieved the purpose of improving the quality of experiments.
Through the introduction of scientific issues,and in combination with the marine training features of Harbin Institute of Technology, the genes of cold-adapted enzymes from marine microorganisms were cloned and the bioinformatics were analyzed in the comprehensive biology experiment. Then the structure characteristics of cold-adapted enzymes were also analyzed using homology modeling. The recombinant enzyme was purified using affinity chromatography. Besides,the genetic engineering products and identify the purity were analyzed by SDS-PAGE. The recombinant enzyme activities were also determined. The comprehensive biology experiment could make students a deep understanding of the research on the genes of cold-adapted enzymes from marine microorganisms,improved their potential for innovation and experimental operation skills,and then achieved the purpose of improving the quality of experiments.
引文
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[15] Humphrey W,Dalke A,Schulten K. VMD:visual molecular dynamics[J]. Journal of Molecular Graphics,1996(1):33-38.
[16] Shi Y L,Wang Q F,Hou Y H,et al. Molecular cloning,expression and enzymatic characterization of glutathione S-transferase from Antarctic sea-ice bacteria Pseudoalteromonas sp. ANT506[J].Microbiological Research,2014,169(2-3):179-184.
[2]粟桂娇,莫祺红,阎欲晓,等.生物工程综合实验独立设课及教学改革[J].实验室研究与探索,2008,27(7):107-110.
[3]李艳红,葛风伟,赵惠新.生物工程专业综合性设计性实验课程教学改革探索[J].新疆师范大学学报(自然科学版),2017,36(3):93-96.
[4]郑权,赵峰,荣龙,等.建立设计性生物工程综合实验培养大学生科技创新能力[J].实验室研究与探索,2012,31(11):121-123.
[5]李江华,房峻,郑飞云,等.生物工程综合实验教学创新体系的构建[J].实验室研究与探索,2009,28(4):222-223.
[6]张立奎,周晓见,王宏归,等.海洋生物基因工程学教学的探索与实践[J].教育教学论坛,2013(48):196-198.
[7] Brown T A.基因克隆和DNA分析[M]. 5版.魏群译.北京:高等教育出版社,2007:28-40.
[8] Sheridan R M,Bentley D L. Selectable one-step PCR-mediated integration of a degron for rapid depletion of endogenous human proteins[J]. Biotechniques,2018,60(2):69-74.
[9] Artimo P,Jonnalagedda M,Arnold K,et al. ExPASy:SIB bioinformatics resource portal[J]. Nucleic Acids Research,2012,40(Web Server issue):W597-W603.
[10] Haider N,Sturm-Ramirez K,Khan S U,et al. Unusually high mortality in waterfowl caused by highly pathogenic avian influenza A(H5N1)in Bangladesh[J]. Transboundary&Emerging Diseases,2017,64(1):144-156.
[11] Kumar S,Stecher G,Tamura K. MEGA 7:Molecular evolutionary genetics analysis version 7. 0 for bigger datasets[J]. Molecular Biology&Evolution,2016,33(7):1870-1874.
[12] Krieger E,Nabuurs S B,Vriend G. Homology modeling[J].Methods of Biochemical Analysis,2003(44):509-523.
[13] Siddiqui K S,Williams T J,Wilkins D,et al. Psychrophiles[J].Annual Review of Earth&Planetary Sciences,2013,41(1):87-115.
[14] Tina K G,Bhadra R,Srinivasan N. PIC:Protein interactions calculator[J]. Nucleic Acids Research,2007,35(Web Server issue):W473-W476.
[15] Humphrey W,Dalke A,Schulten K. VMD:visual molecular dynamics[J]. Journal of Molecular Graphics,1996(1):33-38.
[16] Shi Y L,Wang Q F,Hou Y H,et al. Molecular cloning,expression and enzymatic characterization of glutathione S-transferase from Antarctic sea-ice bacteria Pseudoalteromonas sp. ANT506[J].Microbiological Research,2014,169(2-3):179-184.