Unlock instant, AI-driven research and patent intelligence for your innovation.

High temperature resistant ribosomal protein L-15 gene and its polypeptide encoded by it and preparing process

A ribosomal protein, heat-resistant technology, applied in the field of mutation or genetic engineering, which can solve problems such as lack of similarity

Inactive Publication Date: 2005-04-27
HUADA GENE RES & DEV CENT HANGZHOU
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

EC45 encodes 204 amino acids and it shares 100% similarity with ribosomal protein L15 in the open reading frame but no similarity in the 5' and 3' UTR sequences

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • High temperature resistant ribosomal protein L-15 gene and its polypeptide encoded by it and preparing process
  • High temperature resistant ribosomal protein L-15 gene and its polypeptide encoded by it and preparing process
  • High temperature resistant ribosomal protein L-15 gene and its polypeptide encoded by it and preparing process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Construction of a sequencing library

[0035] The sequencing library was constructed using the genome-wide shotgun method. Firstly, Tengchong thermophilic anaerobic bacteria were cultured according to (Yanfen Xue, 2000) modified MB medium (Balch et al., 1979), bacteria were collected according to Marmur (1961) method, and total DNA was extracted. In order to ensure the randomness of sequencing library construction and avoid the problem of hot spots of breakage to the greatest extent, a variety of methods and principles of library construction under different conditions were adopted. Firstly, physical shearing method (including ultrasonic method and shearing with Hydroshear Machine) was used, and then AluI was selected for random partial enzyme digestion according to the genome characteristics of the bacteria. Different intensities were used to process samples during physical shearing, and samples were processed by setting enzyme gradients during enzyme diges...

Embodiment 2

[0036] Example 2: Genome Sequencing

[0037] When sequencing the genome of thermophilic anaerobic bacteria in Tengchong, two automatic sequencers were mainly used: ABI377 and MegaBACE 1000. These two sequencers use the principle of electrophoresis for sequencing (see Figure 2), and can complete 96 samples each time. ABI377 is a product of PE Company, which is a kind of ABI series. It belongs to the slab gel electrophoresis sequencer. MegaBACE 1000 is a product of Pharmacia, which belongs to capillary gel electrophoresis sequencer.

Embodiment 3

[0038] Example 3: Basecalling and sequencing quality monitoring

[0039] The so-called basecalling refers to the process of obtaining the correct base sequence from the raw data file obtained on the sequencer. Since the sequencer obtains the intensity change traces (traces) of light of different wavelengths corresponding to the four bases A, T, G, and C, it is necessary to use a computer to adopt a certain algorithm to correctly identify the bases corresponding to the different traces. . We used Phred software (Ewing B, Hillier L, 1998) because its results are more reliable and its output is more convenient for further analysis by other programs in the same software package.

[0040] The algorithm principle of Phred's basecalling is to judge the base type based on the shape, distance, and signal-to-noise ratio of each peak in the trajectory, and at the same time give the credibility information for this base, that is, the sequencing quality of the base. In large-scale sequen...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

A refractory ribosomal protein L15 gene, the polypeptide coded by it and its preparing process are disclosed. On the basis of full-genom sequencing and analyzing of a thermophilic anaerobe, the refractory ribosomal protein L15 gene is cloned and separated. It can be used to prepare the the transgenic microbes or animal or plant for producing the said gene and recovering the enzyme coded by the said gene. The amino acid sequence for the polypeptide with activity of the said gene, the function-equal object, and a process for preparing, separating and purifying the said polypeptide are also disclosed.

Description

technical field [0001] The present invention relates to mutation or genetic engineering, in particular to a high-temperature resistant ribosomal protein L15 gene sequence, encoded polypeptide and a preparation method. Background technique [0002] Ribosomes are made up of two subunits, one larger and one smaller. When the magnesium ion concentration is 10mmol / L, the large and small subunits aggregate, and when the magnesium ion concentration drops to 0.1mmol / L, they depolymerize again. The large 50S subunit of the prokaryotic ribosome contains 34 proteins and one molecule each of 5S and 23S rRNA. The 60S large subunit of eukaryotic ribosome contains more than 50 kinds of proteins and one molecule of 5S and 28S rRNA. Ribosomal protein L15 is a protein in the large ribosomal subunit, which belongs to the L15P family of ribosomal proteins, and its gene is rplO. L15 directly binds to 23S rRNA in prokaryotic cells and can also form a complex with acidic...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/00C07K14/435C12N15/12C12N15/63C12N15/64C12P21/02
Inventor 杨焕明包其郁林霞胡松年邓亚君
Owner HUADA GENE RES & DEV CENT HANGZHOU