English
Lizogen bakteriyalar xromosomalarida integratsiyalashgan profaglarga ega bo'lgan va lizissiz normal o'sishi va ko'payishi mumkin bo'lgan bakteriyalarga ishora qiladi. Engil bakteriofaglar transduksiya, gen muhandisligi vektorlari va molekulyar biologiya sohalarida keng qo'llanilgan. hujayra kulturasi bakteriyalarning lizogenligini tekshirish uchun ishlatilishi mumkin. Maxsus ishlash usuli quyidagicha:
1. Lizogen bakteriyalar madaniyati: LB qiyalik bilan faollashtirilgan E. coli 225 (l) ni 20 ml LB kulturali eritmasi bo'lgan 250 ml hujayra kulturasiga soling, 30 ° C da 16 soat davomida chayqatib eking, so'ngra undan 2 ml bakterial suspenziya oling va uni boshqasiga ulang, 20 ml LB bulondan iborat 250 ml Erlenmeyer kolbasida 37°C da 2 soat chayqatib, logarifmik fazaga qadar kultura qiling.
2. Erkin fagni istisno qiling: agar tekshiriladigan shtamm Bacillus bo'lsa, uni spora suspenziyasiga aylantirish mumkin, so'ngra lizogen bakteriyalar yuzasida adsorbsiyalanishi va erkin bo'lishi mumkin bo'lgan fagni o'ldirish uchun 80 ° C da 10 daqiqa davomida davolash mumkin; agar tekshirilishi kerak bo'lgan shtamm Bacillus bo'lmasa, sirtdan adsorbsiyalangan va erkin fagni olib tashlash uchun log fazali lizogen hujayralarini yuvish uchun fag yoki 0,2% natriy sitrat eritmasi bilan tayyorlangan antiserumdan foydalaning. 10 daqiqa davomida 4000 r/min santrifüjdan so'ng, supernatant ikki qatlamli plastinkani aniqlash uchun sezgir indikator bakteriyalar bilan qo'shilishi mumkin, bu namunaning erkin fagini va lizogen bakteriyalarni adsorbsiya qiluvchi fagni tekshirish uchun ishlatiladi. Santrifugadan keyin fag Hujayralar induktsiya qilinadi.
3. Lizogen bakteriyalarni qo'zg'atish: santrifüjdan so'ng to'plangan bakteriyalar ikki marta steril oddiy sho'r suv bilan yuviladi va oddiy fiziologik eritma yoki 100 mmol / l Tris-HCL buferi (pH7.0) bilan yakuniy 1011 hujayra/ml kontsentratsiyasiga keltiriladi, bakterial suspenziya uchun qo'shing. Har bir idishga 5 ml bakterial suspenziya, 30 Vt UV chiroq bilan 30 sm masofada 30 soniya davomida nurlantiring, darhol 5 ml ikki marta konsentratsiyali LB kultura eritmasini qo'shing, yaxshilab aralashtiring va qorong'i joyda 37 ° C da 2 soat davomida inkubatsiya qiling.
4. Lizogen shtammlarni tekshirish: Ikki qavatli agar usuliga muvofiq ishlang, yuqoridagi induktsiyalangan lizatga bir necha tomchi xloroform qo'shing va 10 marta suyultirish usuli bilan mos ravishda suyultiring. Suyultirishning uchta suyultirilishini tanlagandan so'ng, har bir suyultirish uchun 0,3 ml oling. 0,2 ml logarifmik fazaga sezgir E. coli 226 bakteriya suspenziyasi bilan yaxshilab aralashtiriladi, so'ngra eritilgan va 45°C ga sovutilgan LB yarim tana muhitini qo'shing, darhol aralashtiring va keyin uni LB qattiq muhit plastinkasiga quying. Qattiqlashgandan so'ng, 37 ℃ da 6 soat davomida inkubatsiya qiling va kuzating. Agar o'stirilgan plastinkada ko'p miqdordagi blyashka paydo bo'lsa, bu sinovdan o'tgan shtammning lizogen ekanligini isbotlaydi.
Bakteriyalarning lizogen xususiyatiga ega yoki yo'qligini tekshirish uchun yuqoridagi amallarni bajaring. Sinov natijalariga ta'sir qiladigan noto'g'ri ishlashga yo'l qo'ymaslik uchun operatsiya vaqtida jarayonning sterilligiga e'tibor bering.
The FAI climbed 5.9 percent year-on-year in the first 11 months of 2018, quickening from the 5.7-percent growth in Jan-Oct, the National Bureau of Statistics (NBS) said Friday in an online statement.
The key indicator of investment, dubbed a major growth driver, hit the bottom in August and has since started to rebound steadily.
In the face of emerging economic challenges home and abroad, China has stepped up efforts to stabilize investment, in particular rolling out measures to motivate private investors and channel funds into infrastructure.
Friday's data showed private investment, accounting for more than 60 percent of the total FAI, expanded by a brisk 8.7 percent.
NBS spokesperson Mao Shengyong said funds into weak economic links registered rapid increases as investment in environmental protection and agriculture jumped 42 percent and 12.5 percent respectively, much faster than the average.
In breakdown, investment in high-tech and equipment manufacturing remained vigorous with 16.1-percent and 11.6-percent increases respectively in the first 11 months. Infrastructure investment gained 3.7 percent, staying flat. Investment in property development rose 9.7 percent, also unchanged.