| 1. | The pcr product was cloned into pmd18 - t vector . the positive recombinant clone was identified by pcr and endonuclease digest
提取重组质粒经pcr鉴定和酶切鉴定后，对插入片段进行序列测定及分析。 |
| 2. | Recombinant cloning vector of pgem - hc and pgem - ha were constructed by ligating the hc and ha genes into pgem t east vector and transformating into jm109 . 3
分别将目的基因hypoderminc和hypodermina与克隆载体( pgemteasy )连接并转化到宿主菌jm109中，构建了重组克隆载体pgem - hc和pgem - ha 。 |
| 3. | Then the linked products were transformed into the high competent cell of e . coli dh5a . based on - complementation of the detective - galactosidase , positive recombinant clone were screened from x - gal plate
从引物和基因序列的比对分析结果看， zhyf006序列与上下游引物的配对比例分别为s4 |
| 4. | In this experiment , ri gene was amplified by pcr method from recombinant cloning vector pt7 - ri , which had been constructed by dr . yu xiu - ping in this laboratory and sequenced by takara biotechnology ( dalian ) co . , ltd .
于秀平博士已经构建了ri的克隆载体pt7 - ri ，测序表明与已知人胎盘ri基因的cdna序列完全相同。本文的工作是在此基础之上完成的。 |
| 5. | In this study , we designed a pair of primers based on the sequence of the upstream and downstream of chicken il - 2 gene . about 600 bp chicken il - 2 cdna fragment was cloned from cona - stimulated chicken splenocytes by reverse transcription - polymerase chain reaction ( rt - pcr ) and was subcloned into puc18 vector . recombinant clone was demonstrated by restriction enzyme digestion and dna sequencing . next , we construct recombinant plasmid pproex ? t - il - 2 . the cdna of chicken il - 2 gene was subcloned into bamh i / hind iii sites of vector . the recombinant plasmid pproex ? t - il - 2 was transformed into e . coli dh5a and the bacteria was induced with iptg . it was demonstrated by sds - page and western blot that a 18kda protein which was equal to chicken il - 2 protein in molecular weight was expressed in e . coli dh5a . the expression level was up to 30 % of the total bacterial proteins . the purified protein was used to prepare the antibody against chicken il - 2 protein
经酶切鉴定及dna序列测定，该基因为鸡il - 2基因，其序列与sundick等报道的完全一致。在此基础上，我们把鸡il - 2基因亚克隆到大肠杆菌原核表达载体pproex ~ ( tm ) ht中，构建重组表达质粒并进行确证性序列测定，重组质粒测序结果表明将编码鸡il - 2成熟蛋白的基因正确地插入到原核表达载体pproex ~ ( tm ) ht的目的位点。重组质粒转化受体菌dh5后用iptg于37进行诱导培养， sds - page和westernblot分析显示，表达的鸡il - 2融合蛋白分子量约为18kda ，表达的融合蛋白经薄层扫描发现目的蛋白表达量约占菌体蛋白的30 。 |
| 6. | We sequence the inserted gene fragment of the indentified recombinant clone . the result is : angiostatin gene orf ( open reading frame ) links with the orf in expression vector correctly . but the first base of the codon aaa coding for lys414 in plg kringle 4 domain mutates from a to g which leads lys change to glu
随后取通过上述鉴定的重组克隆菌，对重组子插入片段测序，结果为： as基因开放读码框与表达载体的读码框正确匹配相连，但在其kringle4区相当于编码plg的lys ~ ( 414 )密码子aaa的第一位碱基由a突变为g ，导致相应的氨基酸残基突变为glu 。 |
| 7. | The gene was cloned into puc19 vector and identified with pcr and eco ri + hind iii digestion . then , one of the positive recombinant clone was sequenced and analysed . the result showed that its sequence was 35 % and 32 % identical to equine ifn - and human ifn - respectively , but it only shared 15 % homology with chicken type i ifn
序列分析表明，该基因阅读开放框架由492个核苷酸组成，与马和人ifn -基因序列的同源性分别为35和32 ，与鸡i型干扰素基因的同源性仅为15 ，与国外报道的鸡ifn -基因序列完全一致。 |
| 8. | Using enterobacter cloacae b8 , the mutated strains b8b and b8f , and the recombinant clones pb and pf , we try to sequence the antagonistic - related genes of enterobacter cloacae b8 by subcloning and genome primering system . the acquired sequences were analyzed with blast program to find any homology to sequences deposited in genebank
以广谱拮抗菌阴沟肠杆菌b8菌株和拮抗活性缺失菌株b8b 、 b8f及从b8b和b8f二菌株克隆获得的重组质粒pb 、 pf为基础，对阴沟肠杆菌b8菌株拮抗相关的b和f基因片段进行序列分析。 |
