trpS5 E. coli BioVector®色氨酰-tRNA合成酶温度敏感大肠杆菌突变菌株Tryptophanyl-tRNA Synthetase Temperature-Sensitive Mutant Strain

BioVector® trpS5 E. coli 色氨酰-tRNA合成酶温度敏感突变菌株 / Tryptophanyl-tRNA Synthetase Temperature-Sensitive Mutant Strain 数据说明书

产品名称: BioVector® trpS5 大肠杆菌突变菌株
同义词: BioVector® trpS44; BioVector® KY4040
中文名称: BioVector® trpS5 大肠杆菌色氨酰-tRNA合成酶温度敏感突变菌株
英文名称: BioVector® trpS5 E. coli Tryptophanyl-tRNA Synthetase Temperature-Sensitive Mutant Strain
产品类别: 基因工程菌株；温度敏感突变菌株
菌株类型: 大肠杆菌 (Escherichia coli)
基因型: F-， trpS5 (trpS44)， 其他遗传标志因具体菌株背景而异 (如 tyrT58(AS)， mel-1 等)
相关菌株: BioVector® KY4040 (携带 trpS5 突变)
构建方法: 从色氨酸营养缺陷型 trpS5 突变体的回复子中分离得到，该突变体产生变异的色氨酰-tRNA合成酶
突变基因: trpS5 (也称为 trpS44) — 色氨酰-tRNA合成酶 (Tryptophanyl-tRNA synthetase, TRSase) 结构基因的温度敏感突变
遗传特性:

1. 基因定位: trpS 基因位于大肠杆菌染色体约 94 分钟处，与 thr 基因具有 60-80% 的共转导频率

2. trpR 相互作用: trpS5 与 trpR 基因产物存在相互作用，可筛选获得 trpR 突变体以抑制其温度敏感表型
   表型特征:

3. 温度敏感生长:

   • 在 30-37°C 允许温度下，可在完全培养基中正常生长

   • 在 42-43.5°C 非允许温度下，在完全培养基中生长受阻

   • 在基本培养基中，高温敏感性得到缓解

4. 色氨酸操纵子阻遏温度敏感性:

   • 在 43.5°C 且色氨酸过量（阻遏条件）下，邻氨基苯甲酸合成酶产量比 36°C 时增加 10 倍

   • 色氨酸合成酶形成也表现出类似的温度敏感性，但程度较轻

5. TRSase 热失活: 在 42°C 富培养基中快速失活，但在基本培养基或氯霉素存在下不被失活

6. 部分去阻遏: 在生长限制温度下，即使存在过量色氨酸，仍发生邻氨基苯甲酸合成酶的部分去阻遏

7. 温度敏感回复: 与 trpR 突变（包括琥珀突变）组合时，双突变体 (trpR trpS5) 表现为温度不敏感，TRSase 在高温下不被失活

8. trpR 依赖性: 特殊类型的 trpR 突变体仅在携带 trpS5 (而非 trpS⁺) 且在高温下生长的菌株中表现出色氨酸操纵子的组成型表达
   研究应用:

9. 基因表达调控研究: 研究色氨酸操纵子阻遏机制，尤其是色氨酰-tRNA 合成酶与 trpR 阻遏蛋白的相互作用

10. 温度敏感突变研究: 作为温度敏感突变模型，研究蛋白质热稳定性及其调控机制

11. 氨酰-tRNA 合成酶功能研究: 研究色氨酰-tRNA 合成酶在蛋白质合成和基因表达调控中的多重功能

12. tRNA 误氨基酰化研究: 用于研究 tRNA 与色氨酰-tRNA 合成酶的识别特异性及误氨基酰化抑制

13. 遗传互作研究: 研究 trpS 与 trpR 等基因间的遗传相互作用网络
    培养基: BioVector® LB 培养基（完全培养基）；BioVector® 基本培养基（如 M9 培养基）（生长限制条件下使用）
    培养温度:

• 增殖/允许温度：30-37°C

• 限制/非允许温度：42-43.5°C
  抗性: 根据具体菌株背景而异（此菌株本身无特定抗生素抗性标志，需通过其他遗传标志筛选）
  产品形式: 冻干粉 / 甘油菌 / 斜面培养物
  质控载体: BioVector® pUC19 质粒（用于转化效率验证）
  宿主菌: BioVector® DH5α 化学感受态细胞（用于质粒构建和扩增对照）
  质量控制: 无菌检测：阴性；表型验证：30°C 生长正常，42°C 富培养基中生长受阻；遗传标志验证：根据具体菌株背景确认
  储存与运输: -80°C 或干冰运输与保存（长期）；4°C 短期保存（斜面）
  生物安全等级: 1
  仅供科研使用

English:
BioVector® trpS5 E. coli Tryptophanyl-tRNA Synthetase Temperature-Sensitive Mutant Strain Datasheet

Product Name: BioVector® trpS5 E. coli Mutant Strain
Synonyms: BioVector® trpS44; BioVector® KY4040
Chinese Name: BioVector® trpS5 大肠杆菌色氨酰-tRNA合成酶温度敏感突变菌株
English Name: BioVector® trpS5 E. coli Tryptophanyl-tRNA Synthetase Temperature-Sensitive Mutant Strain
Product Category: Genetically Engineered Strain; Temperature-Sensitive Mutant Strain
Strain Type: Escherichia coli
Genotype: F-, trpS5 (trpS44), other genetic markers vary by specific strain background (e.g., tyrT58(AS), mel-1, etc.)
Related Strain: BioVector® KY4040 (carrying trpS5 mutation)
Derivation: Isolated among revertants of a tryptophan auxotroph, trpS5, which produces an altered tryptophanyl-tRNA synthetase
Mutated Gene: trpS5 (also designated trpS44) — temperature-sensitive mutation in the structural gene for tryptophanyl-tRNA synthetase (TRSase)
Genetic Characteristics:

1. Gene Mapping: The trpS gene maps at approximately 94 minutes on the E. coli chromosome, with 60-80% cotransduction frequency with the thr gene

2. trpR Interaction: trpS5 interacts with the trpR gene product; trpR mutants can be isolated to suppress the temperature-sensitive phenotype
   Phenotypic Characteristics:

3. Temperature-Sensitive Growth:

   • Grows normally in complete medium at permissive temperature (30-37°C)

   • Growth is restricted in complete medium at non-permissive temperature (42-43.5°C)

   • High-temperature sensitivity is alleviated in minimal medium

4. Temperature-Sensitive Tryptophan Operon Repression:

   • At 43.5°C with excess tryptophan (repression conditions), produces 10 times more anthranilate synthetase than at 36°C or lower temperatures

   • Similar, though less striking, temperature-sensitivity observed for tryptophan synthetase formation

5. TRSase Heat Inactivation: Rapidly inactivated during growth at 42°C in rich media, but not in minimal media or in the presence of chloramphenicol

6. Partial Derepression: Partial derepression of anthranilate synthetase formation occurs in the presence of excess tryptophan at growth-restricting temperatures

7. Temperature-Insensitive Revertants: When combined with trpR mutations (including amber mutations), resulting double mutants (trpR trpS5) are temperature-insensitive, and TRSase is not inactivated at high temperature

8. trpR Dependency: A special class of trpR mutants exhibits constitutivity of the tryptophan operon only in strains carrying trpS5 (but not trpS⁺) grown at high temperatures
   Research Applications:

9. Gene Expression Regulation Studies: Study of tryptophan operon repression mechanisms, particularly the interaction between tryptophanyl-tRNA synthetase and trpR repressor protein

10. Temperature-Sensitive Mutant Studies: Model for studying protein thermostability and its regulatory mechanisms

11. Aminoacyl-tRNA Synthetase Function: Investigation of multiple functions of tryptophanyl-tRNA synthetase in protein synthesis and gene expression regulation

12. tRNA Mischarging Studies: Study of tRNA recognition specificity by tryptophanyl-tRNA synthetase and suppression of mischarging

13. Genetic Interaction Studies: Investigation of genetic interaction networks between trpS and trpR and other genes
    Culture Medium: BioVector® LB Medium (complete medium); BioVector® Minimal Medium (e.g., M9 Medium) (for growth restriction conditions)
    Culture Temperature:

• Proliferation/Permissive Temperature: 30-37°C

• Restrictive/Non-Permissive Temperature: 42-43.5°C
  Resistance: Varies by specific strain background (this strain lacks a specific antibiotic resistance marker; selection relies on other genetic markers)
  Product Format: Freeze-dried powder / Glycerol stock / Slant culture
  Control Vector: BioVector® pUC19 Plasmid (for transformation efficiency validation)
  Host Strain: BioVector® DH5α Chemically Competent Cells (for plasmid construction and amplification control)
  Quality Control: Sterility: Negative; Phenotypic Validation: Normal growth at 30°C, growth restriction in rich medium at 42°C; Genetic Marker Validation: Confirmed per specific strain background
  Storage and Shipment: Store and ship at -80°C or on dry ice (long-term); short-term storage at 4°C (slants)
  Biosafety Level: 1
  For Research Use Only

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