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Preconcentration of aluminum and copper by the multi-walled carbon nanotubes
No full text[[abstract]]本研究主要利用近年在科學領域快速成長之熱門趨勢材料之ㄧ-奈米碳管，作為金屬前濃縮的基質，並用火焰式原子吸收光譜儀來偵測少量樣品中低濃度金屬鋁及銅的濃度
特別權力關係理論再修正之探討─ 從司法院釋字第653號解釋反思學術自由與大學自治
Get PDF[[abstract]]司法院大法官會議釋字第653號解釋，指出羈押被告其權利遭受侵害時，有向法院提起訴訟，請求救濟之機會。本號解釋對於具有身分關係的特別權力關係理論，做了重大的修正，影響深遠
UVB 輻射誘發老鼠角質層細胞株氧化壓力傷害及抗氧化物光保護效應之應用探討
No full text[[abstract]]紫外線屬「電磁波」一般簡寫成UV（ultraviolet），是一種物理性的危害因子。人體暴露紫外線輻射後可能造成的傷害，如紅斑、DNA的斷裂及皮膚癌等。而目前有許多研究都趨向於細胞模式中，添加抗氧化物來評估是否對紫外線暴露具有光保護的效果
The Analysis of Thesis and Dissertation in Taiwan on the Topic of Teacher Evaluation for Professional Development
No full textDiagnosis Analysis for Health Care Quality of Family Medicine Department in the Veterans Hospital
No full textPurification and Characterization of Extracellular Chitinase from Chitinibacter tainanensis
Get PDF中文摘要………………………………………………...………………vi
英文摘要………………………………………………….……………viii
縮寫表……………………………………………………………………x
致謝……………………………………………………..……………….xi
一、前言………………………………………………….………………1
1. 幾丁質(chitin)
2. 幾丁聚醣的生合成
3. 幾丁質酶(chitinase)
4. 幾丁質酶的反應機制
5. 轉醣苷活性(Transglycosylation)
6. 幾丁聚醣應用
二、研究目的……………………………………………………………10
三、實驗材料與儀器……………………………………………………11
1. 材料和藥品
2. 實驗儀器
四、 實驗方法………………………………………..………………13
1. 微生物與幾丁質酶的生產
2. 幾丁質水解實驗(Chitin degradation assays)
3. 硫酸銨沉澱(Ammonium sulfate precipitation)
4. 幾丁質酶的純化(Purification of chitinase)
5. 微生物與醱酵槽
6. 蛋白質電泳SDS-PAGE
7. Native-PAGE
8. TCA蛋白質沉澱
9. 硝酸銀染色(Silver statining)
10. 酵素圖譜活性分析(Zymogram assay) SDS-PAGE
11. 酵素圖譜活性分析(Zymogram assay) Native-PAGE
12. 蛋白質濃度測定(Thermo Scientific Pierce BCA Protein Assay)
13. 幾丁質酶平板活性測試 (Cup-plate assays)
14. 幾丁質酶的定量活性分析(Chitinase Assay Kit)
五、 結果………………………………………………………………25
1. 微生物培養生長
2. 微生物幾丁質酶的生產
3. 幾丁質水解實驗
4. 幾丁質酶的硫酸銨沉澱
5. 幾丁質酶的純化
6. 醱酵槽大量表現胞外幾丁質酶
7. 幾丁質酶平板活性測試
六、 討論………………………………………………………………29
七、 結論………………………………………………………………31
八、 圖表………………………………………………………………32
九、 參考文獻…………………………………………………………58[[abstract]]N-乙醯葡萄糖胺（NAG）是天然多醣類幾丁質水解的最終產物，近年來發現其具有許多生物功能。本研究所使用的幾丁質酶(chitinase)是從台南縣稻田土壤中篩選出來的一株細菌所分泌出來的，經菌種鑑定分析為新屬新種，並以發現地命名為Chitinibacter tainanensis。先前Chen等人研究發現該菌可將幾丁質顆粒分解成可溶性的還原糖NAG，將α-chitin轉化成NAG的產率達75%，-chitin可達98%，但未對胞外幾丁質酶做進一步研究。於本研究中，我們記錄Chitinibacter tainanensis於LB培養液中(含1 mg/mL β-chitin)0~20小時的生長曲線，結果發現Chitinibacter tainanensis於14小時菌數達最高峰。利用試管幾丁質水解實驗，比較培養14小時的菌體與培養液的上清液兩者水解幾丁質的能力，發現培養液的上清液有明顯活性，證明了此幾丁質酶為胞外分泌蛋白。再者，經由不同培養液測試有無幾丁質誘導物的添加實驗中，發現Chitinibacter tainanensis都能分泌出幾丁質酶，因此推測此幾丁質酶可能是非誘導性酵素。將培養完後的上清液，先以30 %硫酸銨沉澱過濾雜蛋白後，再以75 %硫酸銨沉澱，此可達到初步純化與濃縮。本實驗中利用疏水性、離子交換樹脂以及分子篩分離純化目標蛋白質，並以SDS-PAGE與酵素圖譜活性法分析幾丁質酶分子量大小與活性位置。實驗結果發現，利用疏水性層析管柱純化後，約於33、66.2及120 kDa分子量處有幾丁質酶活性顯示。在Native-PAGE實驗中發現，經由疏水性層析管柱純化後的蛋白質分佈，集中於較大的分子量上，由此推測Chitinibacter tainanensis的胞外幾丁質酶可能是一群CDFs。利用Superdex 75管柱已純化出約33 kDa分子量的幾丁質酶。在幾丁質酶平板活性實驗中，比較純化前與純化後活性差異，結果顯示純化後的幾丁質分解的範圍明顯大於未純化的上清液粗蛋白。未來，我們將建立胞外幾丁質酶活性分析的定量方法並選殖胞外幾丁質酶基因，此將作為生產重組幾丁質酶的基礎。
N-acetyl glucosamine (NAG) is the final hydrolysis product of a natural polysaccharide, chitin. It has been found to have many biological functions. Previously, Chen et al’s study had found the bacteria, Chitinibacter tainanensis, can degrade chitin particles into soluble reducing sugar, NAG. However, they don’t further investigate the extracellular chitinase. In this study, we recorded 0 ~ 20 hours growth curve of Chitinibacter tainanensis in LB medium (containing 1 mg / mL β-chitin), and found that Chitinibacter tainanensis bacterial reached a growth peak at 14 hours. Comparing the chitinase activity between bacteria pellet and culture supernatant by chitin degradation assay, we found that only the latter has enzymatic activity, which was suggested as an extracellular chitinase. Furthermore, with or without chitin induction, Chitinibacter tainanensis can secrete chitinase, suggested that chitinase may be a non-inductive enzyme. After the culture supernatant was first precipitated with 30% ammonium sulfate to filtrate contaminating proteins, and then with 75% ammonium sulfate precipitation, this can be achieved initial purification and concentration. In this study, we purified target chitinase by hydrophobic, ion-exchange and gel filtration chromatography. SDS-PAGE and zymogram assay had been performed to analyze chitinase activity as well as their molecular weight. Our results showed that they display chitinase activity around molecular weight of 33, 66.2 and 120 kDa after hydrophobic chromatography. In the Native-PAGE experiment, we found that the distribution of chitinases purified from hydrophobic chromatography shift to the larger molecular weight, which indicated that Chitinibacter tainanensis extracellular chitinase may be a group of CDFs. In addition, we have purified the chitinase with the molecular weight of 33 kDa by Superdex 75 chromatography. In the chitinase plate assay, we compare the chitinase activity before and after hydrophobic chromatography. The results showed that the purified chitinase from hydrophobic chromatography range significantly greater than the non-purified supernatant crude protein. In the future, we will establish the quantitative methods of extracellular chitinase analysis. Cloning of extracellular chitinase genes is the future work in this study as the basis of the future production of recombinant enzyme
The effect of medium addition on flow rate of bioaerosol impactor
Get PDF第一章 前言 1
1-1 研究背景 1
1-2 研究目的 2
第二章 文獻回顧 3
2-1 何謂生物氣膠 3
2-2 生物氣膠的採樣原理 4
2-3 生物氣膠採樣器效率 8
2-4 慣性衝擊器 10
第三章 材料與方法 13
3-1 研究步驟 13
3-2 研究材料 14
3.2.1流量率監測計校正 15
3-2-2生物氣膠採樣器 20
3-3 研究方法 23
3-3-1 培養基添加量評估 24
3-3-2 衝擊式生物氣膠採樣器校正 25
3-3-3 模擬培養基添加量並進行高度量測 26
3-3-4 採樣器置入不同培養基添加量進行流量率監測 27
3-3-5 監測流量率與廠商提供流量率截斷粒徑計算 29
第四章 結果與討論 31
4-1 Andersen不同培養基添加量對流量率之影響 31
4-1-1 Andersen 不同培養基添加量流量率監測結果 32
4-1-2 Andersen 監測流量率與廠商提供流量率截斷粒徑計算 35
4-2 Burkard不同培養基添加量對流量率之影響 36
4-2-1 Burkard不同培養基添加量流量率監測結果 36
4-2-2 Burkard 監測流量率與廠商提供流量率截斷粒徑計算 41
4-3 RCS 培養基置入對流量率之影響 43
4-3-1 RCS 有無置入培養基對流量率之影響 43
第五章 結論與建議 46
5-1 結論 46
5-2 建議 46
參考文獻 48
附 錄 51[[abstract]]生物氣膠採樣器在採樣過程中往往無法掌握因阻抗造成之流量率變化，採樣流量率大小不只影響生物氣膠氣動特性，更進而影響截斷粒徑與捕集效率而導致濃度評估之差異。本研究之目的為利用流量率監測計在採樣期間進行流量率監測，以探討不同培養基添加量對於衝擊式生物氣膠採樣器流量率變化與影響。首先選擇三種衝擊式生物氣膠採樣器(Andersen單階、Burkard、RCS)，分別以不同培養基添加量採樣，同時以氣流監測器進行每秒紀錄一筆流量率之監測工作共30秒。後續則以氣流監測器流量率數據，以累積方式獲得採樣期間累積之真實採樣體積，探討不同培養基添加量對流量率的影響。Andersen單階、Burkard及RCS三種衝擊式生物氣膠採樣器採樣過程中，流量率皆會受到培養基之影響。其中Burkard在培養基添加量達到一定臨界值時，則產生流量率大幅降低現象。而RCS則在加入培養基時，發現其流量率立即產生變化，與未添加培養基時有較大差異。針對衝擊式生物氣膠採樣器採樣前流量率校正時，可能因培養基添加量與現場採樣時不一致而造成採樣誤差。因此在採樣前進行流量率校正時，建議應依照實際採樣時之培養基添加量進行採樣流量率校正，或於採樣期間同時進行流量率監測，以確保採樣品質並減少暴露濃度推估之誤差。
The fluctuation in flow rate caused by pressure drop resulting from filled medium during sampling is not predictable. Moreover, aerodynamic characteristic, cut-off size, and collection efficiency as well as concentration evaluation are influenced by flow rate. In this study, flow rate monitoring device was utilized to measure the flow rate data during sampling, and volume used to study the effect of medium on the flow rate in bioaerosols impactors. Three bioaerosol impactors (Single Stage Andersen Impactor, Burkard Impactor, and RCS Impactor) with different amount of medium were tested, and the flow rates were also recorded every second for 30 seconds. The data were then integrated to calculate the real accumulated sampling volume during sampling period to study the effect of medium addition amount on flow rate. It was found that flow rate will be affected by the addition of medium for all three bioaerosol impactors. The real sampling flow rate of Burkard decreased significantly as the amount of medium reached to a threshold level. The flow rate of RCS also decreased dramatically as soon as medium addition. Bioaerosol sampler’s performance and flow rate accuracy could be negatively affected by the amount of added medium. To minimize the concentration estimation error, the flow rate calibration before sampling should be executed while the same amount of medium is added as field study. And real-time flow rate monitoring is also a practicable method to guarantee sampling quality and improve the accuracy of concentration evaluation
Bromelain fibrosis mechanism of inhibition of lung cell research
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授權書
致謝.....................................................Ⅲ
目錄.....................................................Ⅳ
圖目錄...................................................Ⅴ
中文摘要.................................................Ⅵ
Abstract.................................................Ⅶ
文獻回顧..................................................1
前言......................................................6
材料與方法...............................................13
結果.....................................................21
討論.....................................................23
結論.....................................................25
參考文獻.................................................26
圖表.....................................................32[[abstract]]肺炎的病程中會發展成肺纖維化。許多的研究指出，生長因子的表現失衡是造成肺纖維化的主要原因。目前的研究發現鳳梨酵素具有很多功效，例如抗發炎...等的效果。然而，尚未有研究指出鳳梨酵素有抑制肺纖維化的效果。因此，我們研究鳳梨酵素在A549 細胞(人類肺泡上皮細胞)中的角色。然而，第一型乙型轉型生長因子(TGF-β1)是多功能性的細胞激素並且扮演引起肺纖維化的重要角色。因此，本研究以第一型乙型轉型生長因子刺激人類肺泡上皮細胞(A549 cell)進行體外培養，模擬肺纖維化。接著，我們以酵素免疫吸附法探討鳳梨酵素在不同(1,5,10,20,25μg/ml)劑量下，對於細胞是否造成細胞毒性，結果發現A549細胞在低劑量時有增加趨勢，但在高濃度鳳梨酵素存在下其細胞存活率降低。接著，我們再加入TGF-β1 (10ng/ml)刺激後，再加入先前研究之不同細胞毒性下的鳳梨酵素，探討caspase蛋白的改變，結果指出，A549細胞中cytochrome c、caspase-9、caspase-3、caspase-7、PARP的表現量，在單獨加入TGF-β1有明顯上升現象，而加入鳳梨酵素後都有明顯降低趨勢。綜合上述結果顯示鳳梨酵素對於肺纖維化有抑制的功效。
Pneumonia in the course of the disease will develop into pulmonary fibrosis. The current study found that bromelain has many effects, including anti-inflammatory effect. However, no studies have pointed out that the effect of bromelain to inhibit pulmonary fibrosis.Many studies have demonstrated that dysregulation of growth factors might be the maincauses underlying the mechanism of Pulmonary fibrosis.The present study found that bromelain has anti-inflammatory effect.However, no studies have pointed out that the effect of bromelain to inhibit pulmonary fibrosis. Therefore, we studied the role of bromelain in A549 (human alveolar epithelial cells). However, transforming growth factor (TGF)-β1 is a multifunctional cytokine which also plays an important role in progressive lung fibrosis. Therefore, in this study , we use transforming growth factor (TGF)-β1 stimulation human alveolar epithelial cells (A549) were cultured in vitro, to simulation of lung fibrosis. We use ELISA in the bromelain (1,5,10,20,25 μg / ml) dose, cell viability of A549 cells in appropriate doses rise, but at high concentrations, survival wasthere are lower.We added TGF-β1 stimulation before adding the pineapple enzymes to explore the change of the caspase protein and found that caspase-3, caspase-7, caspase-9, PARP expression alone by adding TGF-β1 significantly increased by adding bromelain significantly reduced. These results suggest that the inhibition efficacy of bromelain for pulmonary fibrosis
Physiology Functional Application of γ-Amino Butyric Acid - A Review
No full text目 錄
授權................................................................................................................ii
誌謝...............................................................................................................iii
目錄...............................................................................................................iv
中文摘要.....................................................................................................viii
英文摘要.......................................................................................................ix
一、γ-胺基丁酸(Gama-aminobutyric acid，GABA)簡介..........................1
1-1中樞神經系統...................................................................................2
1-2周邊系統...........................................................................................3
二、GABA的代謝.......................................................................................4
2-1生物合成...........................................................................................4
2-2分解...................................................................................................6
三、GABA受體的分類...............................................................................7
3-1 GABA受體介紹...............................................................................7
3-2 GABAA受體....................................................................................8
3-3 GABAB受體....................................................................................11
3-4 GABAC受體....................................................................................14
四、GABA的生理功能................................................................................16
4-1降低血壓...........................................................................................16
4-2幫助睡眠...........................................................................................18
4-3抗焦慮...............................................................................................22
4-4抗衰老.............................................................................................24
4-5生殖作用.........................................................................................26
4-6皮膚癒合.........................................................................................28
4-7胃腸運動.........................................................................................29
4-8其他功能..........................................................................................31
五、乳酸菌之代謝產物..............................................................................32
5-1乳酸菌產GABA研究....................................................................33
六、結論......................................................................................................34
七、參考文獻..............................................................................................37
表目錄
表一、藥物應用影響V1細胞的反應………………………......................70
表二、藥物應用對老彌猴V1區域之方位和方向選擇性細胞的百分比...71
表三、微生物產出GABA之含量................................................................72
表四、從乳酪篩選出的可產出GABA的乳酸菌........................................73
表五、各種乳酪之GABA含量....................................................................74
表六、各種乳酪的發醱菌酛之GABA含量................................................75
表七、GABA影響生理功能........................................................................76
圖目錄
圖一、γ-胺基丁酸分子結構圖...................................................................77
圖二、GABA生物合成途徑......................................................................78
圖三、乳酸菌代謝途徑.............................................................................79
圖四、GABAA受體之架構.........................................................................80
圖五、GABAA受體上不同藥物之結合位.................................................81
圖六、GABAA受體促效劑及拮抗劑.........................................................82
圖七、GABAB受體之結構圖.....................................................................83
圖八、比較EGF與GABA對皮膚創傷的控制........................................84
圖九、GABA信號分子涵蓋人體各個器官...............................................85[[abstract]]γ-胺基丁酸(Gama-aminobutyric acid，簡稱GABA)是一種非蛋白胺基酸，主要存在人體的腦部，是目前所發現最主要的抑制性神經傳導物質。GABA具有許多生理功效，從相關文獻證實：(1)降血壓：藉由活化GABA的受體導致血管擴張及抑制ACE的活性達到降低血壓的作用；(2)幫助睡眠：活化GABAA受體會增加慢波睡眠及抑制奇異睡眠；(3)抗焦慮：在GABAA的存在下，benzodiazepines使氯離子通道開啟的頻率增加，因而增強GABA的抑制作用；(4)抗衰老：GABA與GABAA受體的促效劑能減少V1細胞老化；(5)生殖作用：GABA藉由使膜電位改變進而影響鈣離子的通透性，造成細胞內鈣離子濃度的上升引發精子的頂體反應；(6)皮膚癒合：有效抑制發炎反應，刺激表皮細胞再生；(7)胃腸運動：GABA可透過其受體來調控胃腸運動；(8)其他功能：作為飼料添加使禽畜提高存活率，維持穩定生長。
在許多相關研究乳酸菌的文獻中發現，乳酸菌的功能除了菌體本身外，還包括代謝產物例如乳酸、抗菌素、多醣等，有些乳酸菌還能產出GABA。
本文整理目前已發表之期刊論文，綜合上述之特殊功效，更加確信GABA是值得開發且廣泛應用的保健素材。
GABA is the chief inhibitory neurotransmitter in the mammalian central nervous system. It plays a role in regulating neuronal excitability throughout the nervous system. Many studies have showed that GABA was involved in many physiological functions. (1) Activated GABA receptor induces vasodilation and angiotensin converting enzyme (ACE) inhibition. The blood vessel relaxation leads to decrease blood pressure. (2) It is well established that activation of GABAA receptors favors sleep. The GABAA receptor-mediated inhibitory processes decrease waking and increase slow-wave sleep and the paradoxical sleep is decreased by agonistic modulators of GABAA receptors. (3) Benzodiazepine-induced conformational changes increase GABA's receptor affinity, and thus increase the frequency of ion channel openings. Chloride ion influx hyperpolarizes GABA neurons and produces GABA's inhibitory interneuronal effects. (4) Adding GABA and GABA agonists improve the function of cells in the visual cortex (area V1) and facilitate visual function in old animals. (5) The activation of GABAA receptor leading to Ca2+ channels opening are involved in progesterone-induced acrosomal exocytosis in mouse spermatozoa. (6)To inhibit the inflammatory response and stimulate the epidermal revive. (7)GABA receptors control gastrointestinal motility. (8) Be supplement feed to make domestic birds and animail health.
Many Lactic acid bacterium (LAB) studies found, these beneficial effects not only came from bacterium itself but also metabolism products, including lactic acid, bacteriocin, polysaccharide and GABA.
The thesis review and summarize these publications and according to these studies, we believed that GABA is worthy to develop and apply in functional food
Bidirectional regulation of γ-tubulin―FAM134C promoter and function of γ-tubulin in neuronal differentiation
No full text論文口試委員審定 ---------------------------------------------------------i
授權書 ------------------------------------------------------------------------ ii
致謝 ---------------------------------------------------------------------------iii
中文摘要 ---------------------------------------------------------------------1
Abstract-----------------------------------------------------------------------3
目錄 ---------------------------------------------------------------------------5
圖文目錄 ---------------------------------------------------------------------7
第一章 前言 8
第一節 神經分化 ------------------------------------------------------8
第二節 微小管 --------------------------------------------------------- 9
第三節 微小管在神經分化的角色----------------------------------10
第四節 轉錄因子NRF-1 ----------------------------------------------12
第五節 研究目標 ------------------------------------------------------13
第二章 材料與方法 14
第一節 細胞培養-------------------------------------------------------14
第二節 建構γ-tubulin表現質體 ------------------------------------14
第三節 建構γ-tubulin啟動子-報導基因表現質體 --------------15
第四節 建構雙向啟動子-報導基因表現質體 -------------------15
第五節 定點突變-------------------------------------------------------19
第六節 基因轉染-------------------------------------------------------22
第七節 啟動子分析----------------------------------------------------23
第八節 Gel electrophoretic mobility shift assay -------------------24
第九節 神經突量測 ---------------------------------------------------27
第十節 統計分析 ------------------------------------------------------27
第三章 結果 28
第一節 NRF-1結合在γ-tubulin基因啟動子區域----------------28
第二節 NRF-1調控γ-tubulin基因啟動子活性--------------------28
第三節 NRF-1在雙向基因啟動子活性之角色 ------------------29
第四節 γ-tubulin參與神經突生長----------------------------------33
第四章 討論 34
第一節 γ-tubulin 基因表現調控與NRF-1的角色----------------34
第二節 NRF-1促進神經突生長部分經由-tubulin蛋白-------- 34
第三節 雙向啟動子調控----------------------------------------------36
第四節 γ-tubulin-FAM134C雙向啟動子調控--------------------37
第五節 未來展望 ------------------------------------------------------39
第五章 結論 40
參考文獻 ---------------------------------------------------------------------41
圖文 ---------------------------------------------------------------------------45
自述 ---------------------------------------------------------------------------58
圖一 Growth cone的結構圖-------------------------------------------45
圖二 γ-tubulin啟動子含可能的NRF-1結合部位片段的EMSA分析-----------------------46
圖三 突變NRF-1結合部位對γ-tubulin啟動子的影響-----------47
圖四 NRF-1調控γ-tubulin啟動子活性-----------------------------48
圖五 γ-tubulin-FAM134C啟動子序列------------------------------49
圖六 Forward與reverse type啟動子活性分析---------------------50
圖七 不同長度片段雙向啟動子活性分析-------------------------- 51
圖八 NRF-1對雙向啟動子活性的影響-----------------------------52
圖九 大量表現γ-tubulin對神經突生長之影響-------------------- 53
圖十 相鄰基因的可能轉錄作用方向--------------------------------55
圖十一 Head-to-Head相鄰基因轉錄作用型態-----------------------56
圖十二 γ-tubulin與FAM134C基因表現調控及功能模式圖------57[[abstract]]微小管動態的組合與分離與細胞分裂、細胞移動以及蛋白質與囊泡的運送有關，並且在這個過程裡須要γ-tubulin形成微小管核化以及誘導神經突的延伸。在發展中的神經系統中，對於神經突、生長、分枝，微小管的形成扮演一個關鍵的作用，可是在神經細胞中γ-tubulin作用的分子機轉還不清楚。根據我們先前的研究，NRF-1這個轉錄因子能調控與神經突生長有關的很多基因蛋白，例如CD47與synapsin I。為了研究神經的細胞型態我們在神經纖瘤維母細胞中大量表現γ-tubulin觀察是否能促進神經突生長。此外先前的研究顯示出NRF-1能調控GPAT與AIRC基因的雙向轉錄作用。有趣的是，根據生物資訊學的工具發現，γ-tubulin 與 FAM134C之間的共同啟動子也包含一個NRF-1結合位，我們為了研究NRF-1在這個位子的結合能力，使用gel electrophoretic mobility shift assay證明NRF-1會結合在該基因啟動子上。研究發現突變NRF-1結合部位可以降低γ-tubulin基因啟動子活性，大量表現NRF-1則可促進該基因啟動子活性，表現競爭型抑制的NRF-1則會抑制。為進一步釐清NRF-1在雙向轉錄調控的角色，我們建構不同長度雙向啟動子報導基因表現載體，發現這些雙向啟動子確實具有轉錄活性，突變雙向啟動子上的NRF-1結合部位時，轉錄活性也會下降。在神經纖維瘤母細胞中大量表現γ-tubulin不會增加具有神經突的細胞比例，但可以促進神經突長度。根據本篇研究，NRF-1可以調控γ-tubulin與FAM134C之間的共同啟動子活性，且γ-tubulin具有促進神經突生長的活性。
Dynamic assembly and disassembly of microtubules associate with cell division, motility, and directional transport of proteins and vesicles, and require γ-tubulin to form microtubule nucleation in these processes as well as induce axonal elongation. In the developing nervous system, microtubule formation plays a key role in neurite outgrowth, elongation, and branching, whereas the molecular mechanisms of γ-tubulin in neurons remain unknown. According to our previous studies, nuclear respiratory factor (NRF)-1 is a transcription factor and regulates many genes associated with neurite outgrowth, such as CD47 and synapsin I. To understand the role of γ-tubulin in neurons, we overexpressed γ-tubulin in neuroblastoma cells to observe cellular morphology, such as neurite outgrowth. In addition, a previous study showed that NRF-1 regulates bidirectional transcription in intergenic promoter of GPAT and AIRC genes. Interestingly, according to the prediction using bioinformatics tools, there also is a NRF-1 site on intergenic promoter between γ-tubulin and FAM134C genes. Therefore, we proposed that NRF-1 may regulate bidirectional transactivation of these two genes. To address this issue, we used gel electrophoretic mobility shift assay to study NRF-1 binding activity in this site and site-directed mutagenesis, wild type and dominant-negative NRF-1 expression and promoter assays to determine regulation of NRF-1 on this bidirectional promoter. We found that NRF-1 binds to this site and mutation of this site on γ-tubulin promoter decreased its promoter activity. Furthermore, overexpression of NRF-1 increased the promoter activity but that of dominant-negative NRF-1 decreased that. To clarify the role of NRF-1 in regulating bidirectional intergenic promoter activity, we constructed bidirectional promoter reporter vectors, including various lengths of wild type and mutant promoters, and found these wild type promoters exerted transcriptional activity but mutation of NRF-1 binding site decreased that. We also overexressed γ-tubulin in neuroblastoma cells and found that γ-tubulin did not affected percentage of neurite-bearing cells but increased neurite length. According to these results, NRF-1 involves in regulating intergenic promoter between γ-tubulin and FAM134C genes and γ-tubulin increases neurite outgrowth