BioVector? trpS5 E. coli 色氨酰-tRNA合成酶溫度敏感突變菌株 / Tryptophanyl-tRNA Synthetase Temperature-Sensitive Mutant Strain 數(shù)據(jù)說明書

產(chǎn)品名稱: BioVector? trpS5 大腸桿菌突變菌株
同義詞: BioVector? trpS44; BioVector? KY4040
中文名稱: BioVector? trpS5 大腸桿菌色氨酰-tRNA合成酶溫度敏感突變菌株
英文名稱: BioVector? trpS5 E. coli Tryptophanyl-tRNA Synthetase Temperature-Sensitive Mutant Strain
產(chǎn)品類別: 基因工程菌株；溫度敏感突變菌株
菌株類型: 大腸桿菌 (Escherichia coli)
基因型: F-， trpS5 (trpS44)， 其他遺傳標(biāo)志因具體菌株背景而異 (如 tyrT58(AS)， mel-1 等)
相關(guān)菌株: BioVector? KY4040 (攜帶 trpS5 突變)
構(gòu)建方法: 從色氨酸營養(yǎng)缺陷型 trpS5 突變體的回復(fù)子中分離得到，該突變體產(chǎn)生變異的色氨酰-tRNA合成酶
突變基因: trpS5 (也稱為 trpS44) — 色氨酰-tRNA合成酶 (Tryptophanyl-tRNA synthetase, TRSase) 結(jié)構(gòu)基因的溫度敏感突變
遺傳特性:

1. 基因定位: trpS 基因位于大腸桿菌染色體約 94 分鐘處，與 thr 基因具有 60-80% 的共轉(zhuǎn)導(dǎo)頻率

2. trpR 相互作用: trpS5 與 trpR 基因產(chǎn)物存在相互作用，可篩選獲得 trpR 突變體以抑制其溫度敏感表型
   表型特征:

3. 溫度敏感生長:

   • 在 30-37°C 允許溫度下，可在完全培養(yǎng)基中正常生長

   • 在 42-43.5°C 非允許溫度下，在完全培養(yǎng)基中生長受阻

   • 在基本培養(yǎng)基中，高溫敏感性得到緩解

4. 色氨酸操縱子阻遏溫度敏感性:

   • 在 43.5°C 且色氨酸過量（阻遏條件）下，鄰氨基苯甲酸合成酶產(chǎn)量比 36°C 時增加 10 倍

   • 色氨酸合成酶形成也表現(xiàn)出類似的溫度敏感性，但程度較輕

5. TRSase 熱失活: 在 42°C 富培養(yǎng)基中快速失活，但在基本培養(yǎng)基或氯霉素存在下不被失活

6. 部分去阻遏: 在生長限制溫度下，即使存在過量色氨酸，仍發(fā)生鄰氨基苯甲酸合成酶的部分去阻遏

7. 溫度敏感回復(fù): 與 trpR 突變（包括琥珀突變）組合時，雙突變體 (trpR trpS5) 表現(xiàn)為溫度不敏感，TRSase 在高溫下不被失活

8. trpR 依賴性: 特殊類型的 trpR 突變體僅在攜帶 trpS5 (而非 trpS?) 且在高溫下生長的菌株中表現(xiàn)出色氨酸操縱子的組成型表達(dá)
   研究應(yīng)用:

9. 基因表達(dá)調(diào)控研究: 研究色氨酸操縱子阻遏機制，尤其是色氨酰-tRNA 合成酶與 trpR 阻遏蛋白的相互作用

10. 溫度敏感突變研究: 作為溫度敏感突變模型，研究蛋白質(zhì)熱穩(wěn)定性及其調(diào)控機制

11. 氨酰-tRNA 合成酶功能研究: 研究色氨酰-tRNA 合成酶在蛋白質(zhì)合成和基因表達(dá)調(diào)控中的多重功能

12. tRNA 誤氨基酰化研究: 用于研究 tRNA 與色氨酰-tRNA 合成酶的識別特異性及誤氨基?；种?/p>

13. 遺傳互作研究: 研究 trpS 與 trpR 等基因間的遺傳相互作用網(wǎng)絡(luò)
    培養(yǎng)基: BioVector? LB 培養(yǎng)基（完全培養(yǎng)基）；BioVector? 基本培養(yǎng)基（如 M9 培養(yǎng)基）（生長限制條件下使用）
    培養(yǎng)溫度:

• 增殖/允許溫度：30-37°C

• 限制/非允許溫度：42-43.5°C
  抗性: 根據(jù)具體菌株背景而異（此菌株本身無特定抗生素抗性標(biāo)志，需通過其他遺傳標(biāo)志篩選）
  產(chǎn)品形式: 凍干粉 / 甘油菌 / 斜面培養(yǎng)物
  質(zhì)控載體: BioVector? pUC19 質(zhì)粒（用于轉(zhuǎn)化效率驗證）
  宿主菌: BioVector? DH5α 化學(xué)感受態(tài)細(xì)胞（用于質(zhì)粒構(gòu)建和擴(kuò)增對照）
  質(zhì)量控制: 無菌檢測：陰性；表型驗證：30°C 生長正常，42°C 富培養(yǎng)基中生長受阻；遺傳標(biāo)志驗證：根據(jù)具體菌株背景確認(rèn)
  儲存與運輸: -80°C 或干冰運輸與保存（長期）；4°C 短期保存（斜面）
  生物安全等級: 1
  僅供科研使用

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 大腸桿菌色氨酰-tRNA合成酶溫度敏感突變菌株
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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