English
Hujayra lizisi hujayra o'tkazuvchanligi va faolligini o'zgartirishning muhim eksperimental usuli hisoblanadi. Bu shuni anglatadiki, hujayra virus bilan zararlangandan so'ng, virus o'z naslini sintez qilish uchun xost hujayrasidagi ozuqa moddalaridan foydalanadi va keyin virus avlodi xost hujayrasini yo'q qiladi va hujayradan tashqariga chiqariladi. Hujayra lizisi tajribalarida hujayra kulturalari keng tarqalgan sarflanadigan materialdir. Hujayra lizisining bosqichlari quyidagicha:
1. Madaniyat muhitini to'kib tashlang va hujayra madaniyati shishasini changni yutish qog'ozga teskari bog'lab qo'ying, shunda changni yutish qog'oz madaniy muhitni o'zlashtiradi (yoki qoldiq madaniyat muhiti shishaning pastki qismiga oqib ketishi uchun shishani bir muddat tik qo'ying. va keyin uni so'rish uchun pipetkadan foydalaning ).
2. Har bir shisha idishga 3 ml 4℃ oldindan sovutilgan PBS (0,01M pH7,2～7,3) qo'shing. Hujayralarni tekis qilib qo'ying va hujayralarni yuvish uchun 1 daqiqa davomida muloyimlik bilan silkiting va keyin yuvish eritmasini tashlang. Yuqoridagi amalni ikki marta takrorlang, kultura suyuqligini yuvish uchun hujayralarni uch marta yuving. PBSni tashlang va kulturani muz ustiga qo'ying.
3. 1 ml lizis buferiga 10ul PMSF (100mM) qo'shing, yaxshilab silkiting va muz ustiga qo'ying. (PMSF lizis eritmasi bilan aralashtirishdan oldin kristallar qolmaguncha chayqatilishi kerak.)
4. Har bir hujayra kolbasiga 400 ul PMSF o'z ichiga olgan lizis buferini qo'shing va 30 daqiqa davomida muz ustida lizing. Hujayralarni to'liq parchalash uchun kolbani tez-tez silkitib turish kerak.
5. Lizisdan so'ng, kultura kolbasining bir tomonidagi hujayralarni qirib tashlash uchun toza qirg'ichdan foydalaning (ta'sir tez bo'lishi kerak), so'ngra hujayra qoldiqlarini va lizatni 1,5 ml santrifüj naychasiga o'tkazish uchun quroldan foydalaning. (Barcha operatsiya iloji boricha muz ustida bajarilishi kerak.)
6. 12000 rpm tezlikda 4°C da 5 minut santrifüj qiling. (Oldindan sovutish uchun sentrifugani oldindan yoqing)
7. Santrifüjlangan supernatantni 0,5 daqiqa davomida santrifüj trubasiga o'tkazing va uni -20 ° C da saqlang.
Hujayra kulturalari nafaqat hujayra lizisi tajribalari uchun, balki laboratoriyada Vero hujayralari kabi o'rta miqyosdagi hujayra va to'qimalar madaniyati uchun ham qo'llaniladi. , HEK 293 hujayralari, CAR-T xujayralari, CHO hujayralari, hasharotlar hujayralari va boshqalar.
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.